Skip to main content
Download PDF

Comparison of the quality of ovarian tissue cryopreservation by conventional slow cryopreservation and vitrification—a systematic review and meta-analysis

Journal of Ovarian Research volume 18, Article number: 62 (2025) Cite this article

Abstract

Background

Ovarian tissue cryopreservation is increasingly applied in patients undergoing gonadotoxic radiotherapy or chemotherapy treatment or other patients who need to preserve their fertility. However, there is currently limited evidence to know which type of ovarian tissue cryopreservation is better. The advantages and disadvantages of conventional slow cryopreservation and vitrification are still controversial. The purpose of this meta-analysis was to analyze the ovarian tissue quality of ovarian tissue cryopreservation by conventional slow cryopreservation and vitrification.

Methods

According to the keywords, Pubmed, Embase, and Cochrane Library were searched for studies to January 2024. Studies comparing the follicular viability of conventional slow cryopreservation versus vitrification were assessed for eligibility. The meta-analysis was performed using Stata software (Version 12.0) and Review Manager (Version 5.2).

Results

A total of 18 studies were included in this meta-analysis. The pooled results of the primary outcomes indicated that there was no difference between the two approaches for follicular viability (RR = 0.96, 95% CI: 0.84–1.09, P = 0.520, I2 = 95.8%, Random-effect), the proportion of intact primordial follicles (RR = 1.01, 95% CI: 0.94–1.09, P = 0.778, I2 = 70.6%, Random-effect). The pooled results of the secondary outcomes indicated that there was no difference between the two approaches for the proportion of DNA fragmented follicles (RR = 1.20, 95% CI: 0.94–1.54, P = 0.151, I2 = 0.0%, Fixed-effect), and the proportion of stromal cells (RR = 0.58, 95% CI: 0.20–1.65, P = 0.303, I2 = 99.7%, Random-effect).

Conclusions

Conventional slow cryopreservation and vitrification appear to provide comparable outcomes. The heterogeneity of the literature prevents us from comparing these two techniques. Further high-quality studies are needed to enhance this statement. This meta-analysis provides limited data which may help clinicians when counselling patients.

Introduction

Ovarian tissue cryopreservation has become a widely adopted and significant method for fertility preservation as an increasing number of women become aware of its benefits [1,2,3,4]. Compared with oocyte cryopreservation, ovarian tissue cryopreservation is the only option for preserving fertility in women undergoing cancer treatment that cannot be delayed or in prepubertal girls for whom mature germ cells are not available [5,6,7,8,9,10]. Additionally, ovarian cryopreservation can also be applied to patients with ovarian benign tumors that require ovarian removal and premature ovarian failure. Advancements in autologous transplantation technology following ovarian cryopreservation are also making significant strides, enabling the restoration of endocrine function [11,12,13,14,15].

Since the birth of the first baby using ovarian tissue cryopreservation in 2004, more than 200 babies have been born worldwide through this technology [16, 17]. There are currently two main cryopreservation methods, respectively conventional slow freezing and vitrification [18, 19]. Before cryopreservation, the ovarian tissue should be obtained by laparoscopy or laparotomy. The ovarian tissue with immature germ cells of one or both ovaries is separated into small pieces about 5 × 5 mm2 with a thickness of 1–3 mm before cryopreservation [20].

Conventional slow freezing uses a controlled-rate freezer to gradually cool ovarian tissue with a lower concentration of cryoprotectant, minimizing ice crystal formation and reducing cell damage. Cryoprotectants help limit intracellular ice but may be cytotoxic. Extending freezing time and lowering cryoprotectant levels further reduce cell damage risks. After freezing, the tissue is stored in cryovials within liquid nitrogen at around − 196 °C [21, 22].

Vitrification rapidly cools ovarian tissue to minimize intracellular ice formation, using higher concentrations of cryoprotectants like Dimethyl sulfoxide (DMSO), glycerol, propylene glycol (PrOH), or ethylene glycol (EG) as osmotic agents. While high concentrations increase cytotoxicity, they reduce tissue stress by limiting ice formation. Minimizing exposure to these agents also lowers cell damage risk. Rapid cooling turns the solution into a glass-like, amorphous state, and the tissue is stored in liquid nitrogen at approximately − 196 °C [23,24,25].

Thawing is a crucial step in cryopreservation, warming frozen tissue back to physiological temperature before transplantation. The ovarian tissue is gradually warmed to prevent intracellular ice crystal formation, using specialized warming devices. Afterward, cryoprotectants are removed, and histological examination assesses tissue quality [26].

Histological examination of ovarian tissue is essential for evaluating the success of ovarian tissue cryopreservation and forms the basis for ovarian transplantation. Follicle viability is the most important aspect in the histological examination of ovarian tissue, serving as a key indicator of fertility [27, 28]. Common methods are hematoxylin-eosin (HE) staining, dead and alive cell detection, etc. Since freezing can increase DNA damage, assessing DNA integrity is also essential and is a critical criterion for evaluating the impact of ovarian tissue cryopreservation [29, 30]. The stromal cells of the ovary are a key component of the ovarian tissue, playing a crucial role in supporting the development and function of ovarian follicles [31]. Therefore, the viability and number of stromal cells in the ovarian tissue cryopreservation also determine the growth of follicles and ovarian function after thawing [32]. After thawing the ovarian tissue, ovarian transplantation is the best way to preserve the patient’s fertility and endocrine function. In addition, there are several other ways to culture immature follicles and then transplant them. The culture methods are in vitro follicle culture, in vitro maturation of immature oocytes, and artificial ovary [33, 34]. The schematic overview of ovarian cryopreservation is shown in Fig. 1.

This article aimed to perform a systematic review and meta-analysis to assess the condition of primary and secondary oocytes, stromal cells, and DNA damage in ovarian tissues following conventional slow freezing and vitrification, providing a quality evaluation.

Fig. 1
figure 1

Schematic overview of ovarian cryopreservation

Full size image

Methods

Search strategy

The study was conducted according to the meta-analysis of observational studies in epidemiology (MOOSE) guidelines. We performed a literature search using the keywords “ovarian tissue”, “ovary”, follicle”, “cryopreservation”, and “vitrification” in various combinations to January 2024 in PubMed, Embase, and Cochrane Library. We conducted a Pubmed search using the following search string: (“ovary“[MeSH Terms] OR “ovarial“[All Fields] OR “ovary“[All Fields] OR “ovaries“[All Fields] OR “ovary s“[All Fields] OR ((“ovarian“[All Fields] OR “ovarians“[All Fields]) AND (“tissue s“[All Fields] OR “tissues“[MeSH Terms] OR “tissues“[All Fields] OR “tissue“[All Fields])) OR (“follicle s“[All Fields] OR “hair follicle“[MeSH Terms] OR (“hair“[All Fields] AND “follicle“[All Fields]) OR “hair follicle“[All Fields] OR “follicles“[All Fields] OR “ovarian follicle“[MeSH Terms] OR (“ovarian“[All Fields] AND “follicle“[All Fields]) OR “ovarian follicle“[All Fields] OR “follicle“[All Fields])) AND (“cryopreservability“[All Fields] OR “cryopreservable“[All Fields] OR “cryopreservant“[All Fields] OR “cryopreservants“[All Fields] OR “cryopreservated“[All Fields] OR “cryopreservation“[MeSH Terms] OR “cryopreservation“[All Fields] OR “cryopreserved“[All Fields] OR “cryopreservations“[All Fields] OR “cryopreservative“[All Fields] OR “cryopreservatives“[All Fields] OR “cryopreserve“[All Fields] OR “cryopreserving“[All Fields]) AND (“vitrificated“[All Fields] OR “vitrification“[MeSH Terms] OR “vitrification“[All Fields]). We conducted an Embase search using the following search string: ((‘ovary’/exp OR ovary OR ovarian) AND (‘tissue’/exp OR tissue) OR ‘follicle’/exp OR follicle) AND (‘cryopreservation’/exp OR cryopreservation) AND (‘vitrification’/exp OR vitrification). We conducted a Cochrane Library search using the following search string: (ovary OR ovarian tissue OR follicle) AND cryopreservation AND vitrification. The search All studies were assessed by two investigators independently and any difference was settled by discussion. Studies in all languages were included.

Study selection criteria and exclusion criteria

Research articles comparing the follicular viability of conventional slow cryopreservation and vitrification were considered appropriate for the analysis. Studies that were not classified as research articles were excluded. And studies focused on other topics or with insufficient data for follicular viability were also excluded.

Outcome measures

Primary outcomes

  • Follicular viability (the proportion of viable follicles in the total follicles).

  • The proportion of intact primordial follicles (the proportion of intact primordial follicles in the total primordial follicles).

Secondary outcomes

  • The proportion of DNA fragmented follicles (the proportion of DNA damage follicles in the total follicles).

  • The proportion of stromal cells (the proportion of intact stromal cells in the total stromal cells).

Data abstraction and quality assessment

The following data were extracted: baseline characteristics including age range, mean age, how patients were found, the reason for exclusion of participants, surgical techniques, the method of follicles quality evaluation, the freezing solution of conventional slow freezing and vitrification, follicular density, follicular viability, the proportion of intact primordial follicles, the proportion of DNA fragmented follicles. The risk of bias in individual studies was assessed by the Funnel plot bias. The Newcastle-Ottawa Scale for evidence-based medicine checklist about each risk of bias item as percentages across all included studies were used to evaluate the methodological quality.

Statistical analysis

This meta-analysis was performed using Stata software (Version 12.0) and Review Manager (Version 5.2). We used the Cochran Q test to evaluate the heterogeneity. Heterogeneity was used to evaluate the percentage of the variation in all studies. Because of inevitable clinical and methodologic diversity, inconsistency (I2) was adopted to quantify the effect of statistical heterogeneity. A value of 0% indicated no observed heterogeneity and a value of more than 50% was considered substantial heterogeneity. If I2 > 50%, the random-effects model was adopted, otherwise, the fixed-effects model was used to pool the results. Review Manager (Version 5.2) was used to do the risk of bias graph. A two-sided P < 0.05 was considered statistically significant.

Results

Study characteristics

The search process is shown in Fig. 2. The number of records identified through PubMed, Embase is 1353. The total number of records is 1394 added the records identified through the Cochrane Library. After screening titles and abstracts, 32 publications remained. In the 32 publications, 5 of them have no comparison group, 2 of them are the analysis of xenotransplantation, and 7 of them have no sufficient data. Finally, 18 publications were selected for this meta-analysis [35,36,37,38,39,40,41,42,43,44,45,46,47,48,49,50,51,52].

Fig. 2
figure 2

Flow diagram describing the research selection progress

Full size image

Details of the 18 publications are shown in Tables 1 and 2. All the included studies are published between 2007 and 2022. Published at most in 2022 is 3 articles. The included studies are from 10 countries, China for 5 studies is the most, and Germany for 3 is the second. The mean age range is nearly 20–40 years old. Laparoscopy surgery is the most common surgical approach 7 publications use laparoscopy surgery to obtain ovarian tissue, and 4 publications use laparoscopy surgery or open surgery to obtain ovarian tissue. All cryopreservation ovarian tissue was analyzed by light microscopy.

Table 1 Main characteristics of the selected researches
Full size table
Table 2 Main method and outcome of the selected researches
Full size table

Primary outcome

Follicular viability

The follicular viability of conventional slow cryopreservation versus vitrification in the 18 publications was analyzed. The pooled results indicated that there was no difference in follicular viability between the two approaches from random effect analysis (RR = 0.96, 95% CI: 0.84–1.09, I2 = 95.8%, P < 0.001 for heterogeneity, P = 0.520 shown in Fig. 3A). S.Silber et al.’s study reported vitrification was associated with significantly higher follicular viability (RR = 0.47, 95% CI: 0.43–0.52) [40]. Other two studies from A. Dalman et al. (RR = 1.07, 95% CI: 1.01–1.12) and S.Zhao et al. (RR = 1.16, 95% CI: 1.08–1.25) reported conventional slow cryopreservation was associated with higher follicular viability [42, 49]. Other studies reported there was no difference in follicular viability between the two approaches.

The proportion of intact primordial follicles

The proportion of intact primordial follicles of conventional slow cryopreservation versus vitrification in the 8 publications was analyzed. The other 10 publications did not report the data of the proportion of intact primordial follicles. The pooled results indicated that there was no difference in the proportion of intact primordial follicles between the two approaches from random effect analysis (RR = 1.01, 95% CI: 0.94–1.09, I2 = 70.6%, P = 0.001 for heterogeneity, P = 0.778 shown in Fig. 3B). S.Zhao et al.’s study reported conventional slow cryopreservation was associated with higher intact primordial follicles (RR = 1.18, 95% CI: 1.09–1.26) [42]. Other studies reported there was no difference in intact primordial follicles between the two approaches.

Fig. 3
figure 3

Forest plot for primary outcome (A) Forest plot for the follicular viability of traditional slow cryopreservation and vitrification (B) Forest plot for the proportion of intact primordial follicles of traditional slow cryopreservation and vitrification

Full size image

Second outcome

The proportion of DNA fragmented follicles

The proportion of DNA fragmented follicles of conventional slow cryopreservation versus vitrification in the 5 publications was analyzed. The other 13 publications did not report the data of the proportion of DNA fragmented follicles. The pooled results indicated that there was no difference in the proportion of DNA fragmented follicles between the two approaches (RR = 1.20, 95% CI: 0.94–1.54, I2 = 0.0%, P = 0.675 for heterogeneity, P = 0.151 shown in Fig. 4A).

The proportion of stromal cells

The proportion of stromal cells of conventional slow cryopreservation versus vitrification in the 3 publications was analyzed. The other 15 publications did not report the data of the proportion of stromal cells. The pooled results indicated that there was no difference in the proportion of stromal cells between the two approaches from random effect analysis (RR = 0.58, 95% CI: 0.20–1.65, I2 = 99.7%, P < 0.001 for heterogeneity, P = 0.303 shown in Fig. 4B). The study from V. Keros et al. (RR = 0.23, 95% CI: 0.22–0.23) and H. J. Chang et al. (RR = 0.79, 95% CI: 0.72–0.86) reported vitrification was associated with significantly higher stromal cells [48, 52]. The other study reported there was no difference in the proportion of stromal cells between the two approaches.

Fig. 4
figure 4

Forest plot for second outcome (A) Forest plot for the proportion of DNA fragmented follicles of traditional slow cryopreservation and vitrification (B) Forest plot for the proportion of stromal cells of traditional slow cryopreservation and vitrification

Full size image

Risk of bias in included studies

Funnel plot bias of the studies included in this meta-analysis was notably symmetrical indicating that there was no obvious bias in publication shown in Fig. 5. According to the Newcastle-Ottawa Scale for evidence-based medicine checklist about each risk of bias item as percentages across all included studies, the risk of bias graph was shown in Fig. 6.

Fig. 5
figure 5

Funnel plot bias of the studies included in this meta-analysis

Full size image
Fig. 6
figure 6

Risk of bias graph presenting authors’ judgments according to Newcastle-Ottawa Scale for evidence-based medicine checklist about each risk of bias item as percentages across all included studies

Full size image

Discussion

The meta-analysis is to compare the quality of ovarian tissue cryopreservation by conventional slow cryopreservation and vitrification. A total of 18 articles met the criteria were included. No more articles were included due to that ovarian tissue cryopreservation is not a routine method for every reproductive center, more centers use oocyte cryopreservation to preserve patients’ fertility [53, 54]. First, due to complex ethical methods, ovarian tissue cryopreservation has not yet become a routine method recognized by the public. Second, oocyte cryopreservation can meet the basic requirements of patients for preserving fertility. The cryopreservation of ovarian tissue aims to better preserve endocrine function, allowing follicles to mature and develop more effectively, thereby more accurately maintaining the patient’s fertility [55,56,57,58]. In recent years, with advancements in science and technology and increased awareness, the number of ovarian tissue cryopreservation has increased gradually [27, 52, 59, 60]. There is also controversy about the method of ovarian tissue cryopreservation [61]. Conventional slow cryopreservation can minimize the formation of intracellular ice crystals during the freezing process through gradient cooling. To ensure cell viability, this method has been widely used since the development of ovarian cryopreservation and has achieved good results. The first ovarian transplant birth was the application of conventional slow cryopreservation [62]. However, conventional slow cryopreservation takes a long time and leads to cells death, especially increased [63]. Recently, vitrification has been gradually popular. The vitrification can freeze tissues more quickly though speeding up the freezing by increasing the concentration of cryoprotectant. However, vitrification will lead to the loss of cytoskeletal elements and is more likely to lead to cellular ischemia [64, 65]. As whether the newer freezing method can replace the conventional slow cryopreservation method has become a topic of significant consideration.

The primary outcomes

Follicular viability

As we all know, in ovarian cryopreservation, the most important evaluation criterion is follicular viability [28, 66, 67]. Follicular viability is the basis for determining the patient’s subsequent fertility and is also an important indicator for evaluating ovarian cryopreservation methods. Comparing follicular viability between the two cryopreservation methods, we find that S. Silber et al.‘s study reported significantly higher follicular viability with vitrification, with viability rates of 41.7% for slow freeze-cryopreserved tissue and 89.1% for vitrified tissue [40]. This is quite different from other studies. Without subjective bias, we analyze the reasons for the difference of follicular viability in conventional slow cryopreservation and vitrification. First, in this article, the freezing solution of conventional slow cryopreservation did not contain human serum albumin (HSA) a cryoprotectant that helps protect cells by reducing cellular ice crystal formation [68]. The article was published in 2010, and followed the ovarian transplants for up to 5 years, so the method may be used in 2005 or before. The HSA may not be used in those years as cryoprotectant. Recently, HSA has been widely used for ovarian tissue cryopreservation. In addition to reducing the formation of intracellular ice crystals, it can also maintain cell osmotic pressure and enhance cellular antioxidant capacity [69]. Although it is not an essential component of cryoprotectants, with it can reduce cell damage. Second, the follicles quality evaluation method of this article is different from other articles. Most other articles use microscope counting to judge. This article uses propidium iodide (PI) staining and flow cytometry for detection and assessment. Microscopic counting may not effectively observe subtle changes in cell apoptosis, as early-stage apoptotic cells are harder to identify. However, PI staining can accurately identify cell status. Additionally, transmission electron microscopy is used to observe the cells in this study. The result is that most of the stroma cells in the slow freeze–cryopreserved specimen was lysed and their nuclei compressed between dense bundles of extracellular fibers. Stroma cells are very important in the follicle maturation process, so it may be that the damage to stromal cells caused by conventional slow cryopreservation further led to the damage to follicle cells [70, 71].Third, the freezing cryoprotectant of vitrification is 7.5% ethylene glycol (EG) and 7.5% DMSO for 25 min followed by a second equilibration in 20% EG and 20% DMSO for 15 min. The first concentration is lower than other vitrification methods. Additionally, synthetic serum substitute (SSS) was added as a protective agent, which may contribute to better follicular viability. However, A. Dalman et al. and S.Zhao et al. reported conventional slow cryopreservation was associated with higher follicular viability. A. Dalman et al. reported the follicular viability of conventional slow cryopreservation was 97.4% and vitrification was 91.4% [49]. The follicular viability in this study is higher than in other studies. This study uses microscope counting for detection and assessment. There may be some subjective bias. At the same time, vitrification was correlated with more free radicals, and reactive oxygen species (ROS) [72]. Oxidative stress occurs when more ROS produced, which will result in damage to cellular components such as DNA, proteins, and lipids and cause cell damage [73, 74]. Adding catalase to the cryoprotectant may reduce the production of ROS and protect freezing cells [75]. S.Zhao et al. reported the follicular viability of conventional slow cryopreservation was 82.9% and vitrification was 71.3% [42]. The main reason for this difference is that this study used large pieces of human ovarian tissues that is 15 mm × 15 mm × 2 mm which is larger than other studies. The larger ovarian tissue cryopreservation may reduce follicles loss and support follicles growth [76, 77]. The higher density of the vitrification cryoprotectant prevents it from quickly penetrating large tissues, resulting in uneven vitrification of the cellular water throughout the tissue. This may be the reason for conventional slow cryopreservation was associated with higher follicular viability.

The proportion of intact primordial follicles

Primordial follicles are the earliest stage of ovarian follicles, which account for more than 90% of the population of follicles [78, 79]. During ovarian tissue cryopreservation, a higher proportion of intact primordial follicles indicates greater fertility potential [43, 80]. Comparing the proportion of intact primordial follicles between the two cryopreservation methods, S.Zhao et al. reported the proportion of intact primordial follicles of conventional slow cryopreservation was 86.8% and vitrification was 73.8% [42]. As previously mentioned, this study used large pieces of human ovarian tissues, which may lead to differing results between the two methods. Larger tissue samples may benefit from conventional slow cryopreservation not vitrification, which may not penetrate large tissues quickly enough, potentially leading to the formation of intracellular ice crystals that can damage cells. More studies about this supposed need to be verified. Additionally, while the morphology of frozen primordial follicles may appear normal, their subsequent development, fertilization, and the ability to achieve pregnancy and live birth can be influenced by various factors [54, 81]. Therefore, fertility outcomes cannot be solely determined by normal morphology; it represents just the initial step in assessing the efficiency of cryopreservation.

Secondary outcomes

The proportion of DNA fragmented follicles

According to this meta-analysis, there was no difference in the proportion of DNA fragmented follicles between the two approaches. Both methods can lead to cell apoptosis, mainly manifested as DNA damage. This is mainly attributed to the inevitable formation of intracellular ice crystals during the freezing process, despite efforts by cryoprotectants to minimize their formation. The formation of intracellular ice crystals can cause mechanical damage, which can further lead to rupture of cell membranes and other cellular structures. Moreover, it can cause structural damage by causing damage and breakage of DNA strands due to reduced space for cell expansion [82, 83]. In addition, cryoprotectants containing that may cause chemical damage and directly damage DNA [84]. Therefore, reducing DNA damage during the freezing process should also be considered in future freezing methods.

The proportion of stromal cells

Stromal cells are an essential part of ovarian tissue providing structural support for follicles and contributing to the production of extracellular matrix components and hormones. They provide the necessary microenvironment for the growth and development of oocytes and play an important role in the development and maturation of follicles [85, 86]. Research shows that the weakened antioxidant capacity of stromal cells is closely related to ovarian aging [87]. More attention should be paid to the survival status of stromal cells in ovarian tissue cryopreservation. The study of H. J. Chang et al. showed that the proportion of stromal cells of conventional slow cryopreservation was 47.8% and vitrification was 60.7%. Vitrification may reduce damage to stromal cells, possibly because stromal cells are relatively small. During vitrification, they can quickly reach the vitrified state, reducing the formation of intracellular ice crystals, and thereby reducing cell damage [69]. However, it’s important to note that the vitrification method is not standardized, and further studies using consistent vitrification protocols are needed for comprehensive evaluation.

Limitations

This meta-analysis reviewed the comparison of ovarian tissue cryopreservation by conventional slow cryopreservation and vitrification till January 2024. Our results indicate comparable outcomes for ovarian tissue cryopreservation between the two approaches. The main limitation of our study is the significant variation in the methods of conventional slow cryopreservation and vitrification. This variability arises from differences in the composition and ratio of cryoprotectants used. They mainly contain such as EG, DMSO, PrOH, etc. The difference between the different cryoprotectants is not particularly obvious. There is currently no unified regulation. Studies by Meryman indicated that osmotic effects may be the major factors in slow-freezing cell injury [88]. The same group demonstrated that adding neutral solutes as cryoprotective agents (CPAs) can reduce osmotically driven cell injury during ice solidification due to colligative effects [89]. There are two main categories of cryoprotective agents [90, 91]. The first is penetrating cryoprotective agents. DMSO is one of the most effective and widely used cryoprotective agents which is a small, amphipathic molecule that can easily penetrate cell membranes and distribute throughout both intracellular and extracellular environments [92]. During the freezing process, DMSO creates an osmotic gradient that helps dehydrate cells, reducing the amount of free water inside them and thereby decreasing the chances of intracellular ice formation. By controlling the osmotic balance, it also helps prevent osmotic shock during freezing and thawing, which can lead to cell lysis or severe deformation. The concentration of DMSO used in cryopreservation typically ranges from 5 to 10% [93,94,95]. EG is another effective cryoprotective agent that has a similar function to DMSO. The main difference is that EG is a small, water-soluble molecule. The concentration of ethylene glycol used in cryopreservation protocols typically ranges from 10 to 20% [96, 97]. In many studies, a mixture of DMSO and EG is reported to be more effective as the cryoprotectant [93, 98,99,100]. The other cryoprotective agents are non-penetrating cryoprotective agents with sucrose being the primary one. The combination of different cryoprotectants aims to enhance osmotic pressure regulation inside and outside cells, prevent intracellular ice crystal formation, and stabilize cell membranes and proteins. Different researchers utilize varying combinations of cryoprotectants based on their individual research experiences [93, 94, 101, 102]. Moreover, follicular viability, the proportion of intact primordial follicles, the proportion of DNA fragmented in follicles, and the proportion of stromal cells are essential criteria for evaluating ovarian tissue cryopreservation. The clinical pregnancy rate is also very important in ovarian tissue cryopreservation and is the most crucial metric for evaluating female reproductive function. However, under current clinical conditions, experience is relatively limited. Beyond evaluating ovarian cryopreservation technology, the success of ovarian tissue cryopreservation also depends on transplantation surgery techniques, the method of conception, and the physical condition of the female patient, including factors such as pelvic radiation and prior chemotherapy. And until now, the ovarian tissue cryopreservation is considered an experimental procedure. More studies are needed.

Conclusion

In conclusion, this meta-analysis indicated that conventional slow cryopreservation and vitrification appear to provide comparable outcomes of follicular viability, the proportion of intact primordial follicles, the proportion of DNA fragmented follicles and the proportion of stromal cells. However, included studies varied in the use of cryopreservation protocols. Therefore, further studies are needed to determine the optimal method for cryopreservation human ovarian tissue. While there is no significant difference between traditional cryopreservation and vitrification regarding ovarian tissue anatomy, each method has its own advantages and is suitable for different clinical scenarios. The advantages of traditional cryopreservation include its mature technology and relatively standardized procedures. On the other hand, vitrification offers benefits such as fast freezing speed, simplicity of operation, and high flexibility. And until now, the ovarian tissue cryopreservation is considered an experimental procedure. The choice of method can be tailored to meet specific clinical needs.

Data availability

No datasets were generated or analysed during the current study.

Abbreviations

DMSO:

Dimethyl sulfoxide

PrOH:

Glycerol, propylene glycol

EG:

Ethylene glycol

HAS:

Human serum albumin

SSS:

Synthetic serum substitute

ROS:

Reactive oxygen species

References

  1. Dolmans MM, Donnez J, Cacciottola L. Fertility preservation: the challenge of freezing and transplanting ovarian tissue. Trends Mol Med. 2021;27(8):777–91.

    Article  CAS  PubMed  Google Scholar 

  2. Donnez J, Dolmans MM. Fertility preservation in men and women: where are we in 2021? Are we rising to the challenge? Fertil Steril. 2021;115(5):1089–90.

    Article  PubMed  Google Scholar 

  3. Yding Andersen C, Mamsen LS, Kristensen SG. FERTILITY PRESERVATION: freezing of ovarian tissue and clinical opportunities. Reproduction. 2019;158(5):F27–34.

    Article  CAS  PubMed  Google Scholar 

  4. Schallmoser A, Einenkel R, Färber C, Hüren V, Pougin A, Emrich N, et al. Cryostorage of human ovarian tissue: evaluating the storage and disposal pattern over a 22-year period in 2475 patients. Reprod Biomed Online. 2023;47(3):103239.

    Article  PubMed  Google Scholar 

  5. Duffin K, Howie R, Kelsey TW, Wallace HB, Anderson RA. Long-term follow-up to assess criteria for ovarian tissue cryopreservation for fertility preservation in young women and girls with cancer. Hum Reprod. 2023;38(6):1076–85.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Dolmans MM, Donnez J. Fertility preservation in women for medical and social reasons: Oocytes vs ovarian tissue. Best practice & research Clinical obstetrics & gynaecology. 2021;70:63–80.

  7. Ruan X, Cheng J, Du J, Jin F, Gu M, Li Y, et al. Analysis of fertility preservation by ovarian tissue cryopreservation in Pediatric Children in China. Front Endocrinol (Lausanne). 2022;13:930786.

    Article  PubMed  Google Scholar 

  8. Rives N, Courbière B, Almont T, Kassab D, Berger C, Grynberg M, et al. What should be done in terms of fertility preservation for patients with cancer? The French 2021 guidelines. Eur J Cancer. 2022;173:146–66.

    Article  PubMed  Google Scholar 

  9. Fabbri R, Vicenti R, Magnani V, Paradisi R, Lima M, De Meis L, et al. Ovarian tissue cryopreservation and transplantation: 20 years experience in Bologna University. Front Endocrinol (Lausanne). 2022;13:1035109.

    Article  PubMed  Google Scholar 

  10. Newton H, Fisher J, Arnold JR, Pegg DE, Faddy MJ, Gosden RG. Permeation of human ovarian tissue with cryoprotective agents in preparation for cryopreservation. Hum Reprod. 1998;13(2):376–80.

    Article  CAS  PubMed  Google Scholar 

  11. Khattak H, Malhas R, Craciunas L, Afifi Y, Amorim CA, Fishel S, et al. Fresh and cryopreserved ovarian tissue transplantation for preserving reproductive and endocrine function: a systematic review and individual patient data meta-analysis. Hum Reprod Update. 2022;28(3):400–16.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Blumenfeld Z. Ovarian tissue transplantation and leukemia. Fertil Steril. 2018;109(1):69–70.

    Article  PubMed  Google Scholar 

  13. Silber SJ, DeRosa M, Goldsmith S, Fan Y, Castleman L, Melnick J. Cryopreservation and transplantation of ovarian tissue: results from one center in the USA. J Assist Reprod Genet. 2018;35(12):2205–13.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Isachenko V, Morgenstern B, Todorov P, Isachenko E, Mallmann P, Hanstein B, et al. Patient with ovarian insufficiency: baby born after anticancer therapy and re-transplantation of cryopreserved ovarian tissue. J Ovarian Res. 2020;13(1):118.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Oktay K, Karlikaya G. Ovarian function after transplantation of frozen, banked autologous ovarian tissue. N Engl J Med. 2000;342(25):1919.

    Article  CAS  PubMed  Google Scholar 

  16. Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;364(9443):1405–10.

    Article  CAS  PubMed  Google Scholar 

  17. Dolmans MM, von Wolff M, Poirot C, Diaz-Garcia C, Cacciottola L, Boissel N, et al. Transplantation of cryopreserved ovarian tissue in a series of 285 women: a review of five leading European centers. Fertil Steril. 2021;115(5):1102–15.

    Article  CAS  PubMed  Google Scholar 

  18. Rienzi L, Gracia C, Maggiulli R, LaBarbera AR, Kaser DJ, Ubaldi FM, et al. Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance. Hum Reprod Update. 2017;23(2):139–55.

    CAS  PubMed  Google Scholar 

  19. Jaeger P, Fournier C, Santamaria C, Fraison E, Morel-Journel N, Benchaib M, et al. Human ovarian cryopreservation: vitrification versus slow freezing from histology to gene expression. Hum Fertil (Camb). 2023;26(5):1099–107.

    Article  CAS  PubMed  Google Scholar 

  20. Meirow D, Baum M, Yaron R, Levron J, Hardan I, Schiff E, et al. Ovarian tissue cryopreservation in hematologic malignancy: ten years’ experience. Leuk Lymphoma. 2007;48(8):1569–76.

    Article  PubMed  Google Scholar 

  21. Isachenko V, Isachenko E, Reinsberg J, Montag M, Braun F, van der Ven H. Cryopreservation of human ovarian tissue: effect of spontaneous and initiated ice formation. Reprod Biomed Online. 2008;16(3):336–45.

    Article  PubMed  Google Scholar 

  22. Sanfilippo S, Canis M, Romero S, Sion B, Déchelotte P, Pouly JL, et al. Quality and functionality of human ovarian tissue after cryopreservation using an original slow freezing procedure. J Assist Reprod Genet. 2013;30(1):25–34.

    Article  PubMed  Google Scholar 

  23. Isachenko V, Montag M, Isachenko E, van der Ven K, Dorn C, Roesing B, et al. Effective method for in-vitro culture of cryopreserved human ovarian tissue. Reprod Biomed Online. 2006;13(2):228–34.

    Article  PubMed  Google Scholar 

  24. Isachenko E, Isachenko V, Nawroth F, Rahimi G, Kreienberg R, Reinsberg J, et al. Human ovarian tissue preservation: is vitrification acceptable method for assisted reproduction? Cryo Lett. 2008;29(4):301–14.

    CAS  Google Scholar 

  25. Martínez-Burgos M, Herrero L, Megías D, Salvanes R, Montoya MC, Cobo AC, et al. Vitrification versus slow freezing of oocytes: effects on morphologic appearance, meiotic spindle configuration, and DNA damage. Fertil Steril. 2011;95(1):374–7.

    Article  PubMed  Google Scholar 

  26. Isachenko V, Dittrich R, Keck G, Isachenko E, Rahimi G, van der Ven H, et al. Cryopreservation of ovarian tissue: detailed description of methods for transport, freezing and thawing. Geburtshilfe Frauenheilkd. 2012;72(10):927–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Isachenko V, Orth I, Isachenko E, Mallmann P, Peters D, Schmidt T, et al. Viability of human ovarian tissue confirmed 5 years after freezing with spontaneous ice-formation by autografting and chorio-allantoic membrane culture. Cryobiology. 2013;66(3):233–8.

    Article  PubMed  Google Scholar 

  28. Isachenko V, Isachenko E, Kreienberg R, Woriedh M, Weiss J. Human ovarian tissue cryopreservation: quality of follicles as a criteria of effectiveness. Reprod Biomed Online. 2010;20(4):441–2.

    Article  CAS  PubMed  Google Scholar 

  29. Kim B, Ryu KJ, Lee S, Kim T. Changes in telomere length and senescence markers during human ovarian tissue cryopreservation. Sci Rep. 2021;11(1):2238.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Noda M, Ma Y, Yoshikawa Y, Imanaka T, Mori T, Furuta M, et al. A single-molecule assessment of the protective effect of DMSO against DNA double-strand breaks induced by photo-and γ-ray-irradiation, and freezing. Sci Rep. 2017;7(1):8557.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Shen L, Liu J, Luo A, Wang S. The stromal microenvironment and ovarian aging: mechanisms and therapeutic opportunities. J Ovarian Res. 2023;16(1):237.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Xia X, Yin T, Yan J, Yan L, Jin C, Lu C, et al. Mesenchymal stem cells enhance angiogenesis and follicle survival in human cryopreserved ovarian cortex transplantation. Cell Transpl. 2015;24(10):1999–2010.

    Article  Google Scholar 

  33. Cho E, Kim YY, Noh K, Ku SY. A new possibility in fertility preservation: the artificial ovary. J Tissue Eng Regen Med. 2019;13(8):1294–315.

    Article  CAS  PubMed  Google Scholar 

  34. Telfer EE, Fauser BC. Important steps towards materializing the dream of developing an artificial ovary. Reprod Biomed Online. 2016;33(3):333–4.

    Article  PubMed  Google Scholar 

  35. Borrás A, Manau D, Fabregues F, Peralta S, Calafell JM, Casals G, et al. Comparison between slow freezing and vitrification of ovarian tissue cryopreservation in assigned female at birth transgender people receiving testosterone therapy: data on histological and viability parameters. J Assist Reprod Genet. 2022;39(2):527–41.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Galbinski S, Kowalewski LS, Grigolo GB, da Silva LR, Jiménez MF, Krause M, et al. Comparison between two cryopreservation techniques of human ovarian cortex: morphological aspects and the heat shock response (HSR). Cell Stress Chaperones. 2021;27(2):97–106.

    Article  PubMed  Google Scholar 

  37. Sugishita Y, Taylan E, Kawahara T, Shahmurzada B, Suzuki N, Oktay K. Comparison of open and a novel closed vitrification system with slow freezing for human ovarian tissue cryopreservation. Reproductive sciences (Thousand Oaks. Calif). 2021;38(10):2723–33.

    Google Scholar 

  38. Huang L, Mo Y, Wang W, Li Y, Zhang Q, Yang D. Cryopreservation of human ovarian tissue by solid-surface vitrification. Eur J Obstet Gynecol Reprod Biol. 2008;139(2):193–8.

    Article  CAS  PubMed  Google Scholar 

  39. Nikhil Kumar Tej J, Somoskoi B, Cseh S, Ramos L, Galbinski S, Nacul A, et al. Detailed morphological analysis of Cryoinjury in Human ovarian tissue following vitrification or slow freezing. Reprod Domest Anim = Zuchthygiene. 2022;29(8):2374–81.

    Google Scholar 

  40. Silber S, Kagawa N, Kuwayama M, Gosden R. Duration of fertility after fresh and frozen ovary transplantation. Fertil Steril. 2010;94(6):2191–6.

    Article  PubMed  Google Scholar 

  41. Schallmoser A, Einenkel R, Färber C, Emrich N, John J, Sänger N. The effect of high-throughput vitrification of human ovarian cortex tissue on follicular viability: a promising alternative to conventional slow freezing? Arch Gynecol Obstet. 2023;307(2):591–9.

    Article  PubMed  Google Scholar 

  42. Zhao S, Zhao SQ, Shi Q, Qin XM, Han YL, Chao L, et al. Effects of different freezing methods on the follicular viability of large pieces of human ovarian tissues. Acta Anat Sinica. 2013;44(3):427–32.

    CAS  Google Scholar 

  43. Isachenko V, Lapidus I, Isachenko E, Krivokharchenko A, Kreienberg R, Woriedh M, et al. Human ovarian tissue vitrification versus conventional freezing: morphological, endocrinological, and molecular biological evaluation. Reprod (Cambridge England). 2009;138(2):319–27.

    Article  CAS  Google Scholar 

  44. Wang TR, Yan J, Lu CL, Xia X, Yin TL, Zhi X, et al. Human single follicle growth in vitro from cryopreserved ovarian tissue after slow freezing or vitrification. Hum Reprod (Oxford England). 2016;31(4):763–73.

    Article  CAS  Google Scholar 

  45. Li YX, Sun YP, Tang L, Ma YP. Influence of different cryopreservation schemes on human ovarian tissue activity. Chin J Tissue Eng Res. 2013;17(53):9125–31.

    CAS  Google Scholar 

  46. Li YB, Zhou CQ, Yang GF, Wang Q, Dong Y. Modified vitrification method for cryopreservation of human ovarian tissues. Chin Med J. 2007;120(2):110–4.

    Article  CAS  PubMed  Google Scholar 

  47. Fabbri R, Vicenti R, Macciocca M, Martino NA, Dell’Aquila ME, Pasquinelli G, et al. Morphological, ultrastructural and functional imaging of frozen/thawed and vitrified/warmed human ovarian tissue retrieved from oncological patients. Hum Reprod (Oxford England). 2016;31(8):1838–49.

    Article  CAS  Google Scholar 

  48. Chang HJ, Moon JH, Lee JR, Jee BC, Suh CS, Kim SH. Optimal condition of vitrification method for cryopreservation of human ovarian cortical tissues. J Obstet Gynaecol Res. 2011;37(8):1092–101.

    Article  PubMed  Google Scholar 

  49. Dalman A, Deheshkar Gooneh Farahani NS, Totonchi M, Pirjani R, Ebrahimi B, Rezazadeh Valojerdi M. Slow freezing versus vitrification technique for human ovarian tissue cryopreservation: an evaluation of histological changes, WNT signaling pathway and apoptotic genes expression. Cell Tissue Banking. 2017;79:29–36.

    CAS  Google Scholar 

  50. Klocke S, Bündgen N, Köster F, Eichenlaub-Ritter U, Griesinger G. Slow-freezing versus vitrification for human ovarian tissue cryopreservation. Arch Gynecol Obstet. 2015;291(2):419–26.

    Article  CAS  PubMed  Google Scholar 

  51. Sanfilippo S, Canis M, Dechelotte P, Artonne C, Janny L, Brugnon F. Vitrification versus slow freezing: what is the better method to preserve human ovarian tissue quality? Fertil Steril. 2012;98(3):S130–1.

    Article  Google Scholar 

  52. Keros V, Xella S, Hultenby K, Pettersson K, Sheikhi M, Volpe A, et al. Vitrification versus controlled-rate freezing in cryopreservation of human ovarian tissue. Hum Reprod (Oxford England). 2009;24(7):1670–83.

    Article  CAS  Google Scholar 

  53. Hossay C, Tramacere F, Cacciottola L, Camboni A, Squifflet JL, Donnez J, et al. Follicle outcomes in human ovarian tissue: effect of freezing, culture, and grafting. Fertil Steril. 2023;119(1):135–45.

    Article  PubMed  Google Scholar 

  54. Fabbri R. Cryopreservation of human oocytes and ovarian tissue. Cell Tissue Banking. 2006;7(2):113–22.

    Article  PubMed  Google Scholar 

  55. Bromer JG, Patrizio P. Fertility preservation: the rationale for cryopreservation of the whole ovary. Semin Reprod Med. 2009;27(6):465–71.

    Article  PubMed  Google Scholar 

  56. Silber SJ. Ovary cryopreservation and transplantation for fertility preservation. Mol Hum Reprod. 2012;18(2):59–67.

    Article  CAS  PubMed  Google Scholar 

  57. Amorim CA, Dolmans MM, David A, Jaeger J, Vanacker J, Camboni A et al. Vitrification and xenografting of human ovarian tissue. Fertility and sterility. 2012;98(5):1291-8.e1-2.

  58. Ovarian tissue cryopreservation. A committee opinion. Fertil Steril. 2014;101(5):1237–43.

    Article  Google Scholar 

  59. Rosendahl M, Schmidt KT, Ernst E, Rasmussen PE, Loft A, Byskov AG, et al. Cryopreservation of ovarian tissue for a decade in Denmark: a view of the technique. Reprod Biomed Online. 2011;22(2):162–71.

    Article  PubMed  Google Scholar 

  60. Bastings L, Liebenthron J, Westphal JR, Beerendonk CC, van der Ven H, Meinecke B, et al. Efficacy of ovarian tissue cryopreservation in a major European center. J Assist Reprod Genet. 2014;31(8):1003–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Edgar DH, Gook DA. A critical appraisal of cryopreservation (slow cooling versus vitrification) of human oocytes and embryos. Hum Reprod Update. 2012;18(5):536–54.

    Article  PubMed  Google Scholar 

  62. Maffei S, Hanenberg M, Pennarossa G, Silva JR, Brevini TA, Arav A, et al. Direct comparative analysis of conventional and directional freezing for the cryopreservation of whole ovaries. Fertil Steril. 2013;100(4):1122–31.

    Article  PubMed  Google Scholar 

  63. Zhang JM, Wang HX, Lu XL. Damage of Granusola and Stroma cells exposed to mouse ovarian tissue cryopreservation: potential mechanism of Ovarian Injury. Cryo Lett. 2021;42(1):53–8.

    CAS  Google Scholar 

  64. Wang D, Geng M, Gan D, Han G, Gao G, Xing A, et al. Effect of resveratrol on mouse ovarian vitrification and transplantation. Volume 19. Reproductive biology and endocrinology: RB&E; 2021. p. 54. 1.

    Google Scholar 

  65. Lornage J, Salle B. Ovarian and oocyte cryopreservation. Curr Opin Obst Gynecol. 2007;19(4):390–4.

    Article  Google Scholar 

  66. Marschalek J, Hager M, Wanderer S, Ott J, Frank M, Schneeberger C et al. Different impacts of Cryopreservation in endothelial and epithelial ovarian cells. Int J Mol Sci. 2023;24(15).

  67. Behl S, Joshi VB, Larson NB, Young MC, Bilal M, Walker DL, et al. Vitrification versus slow freezing of human ovarian tissue: a systematic review and meta-analysis of histological outcomes. J Assist Reprod Genet. 2023;40(3):455–64.

    Article  PubMed  Google Scholar 

  68. Hreinsson J, Zhang P, Swahn ML, Hultenby K, Hovatta O. Cryopreservation of follicles in human ovarian cortical tissue. Comparison of serum and human serum albumin in the cryoprotectant solutions. Hum Reprod (Oxford England). 2003;18(11):2420–8.

    Article  CAS  Google Scholar 

  69. Hreinsson JG, Scott JE, Rasmussen C, Swahn ML, Hsueh AJ, Hovatta O. Growth differentiation factor-9 promotes the growth, development, and survival of human ovarian follicles in organ culture. J Clin Endocrinol Metab. 2002;87(1):316–21.

    Article  CAS  PubMed  Google Scholar 

  70. Kinnear HM, Tomaszewski CE, Chang FL, Moravek MB, Xu M, Padmanabhan V, et al. The ovarian stroma as a new frontier. Reprod (Cambridge England). 2020;160(3):R25–39.

    Article  CAS  Google Scholar 

  71. Fienberg R, Cohen RB, A COMPARATIVE HISTOCHEMICAL STUDY OF THE OVARIAN STROMAL LIPID BAND, STROMAL THECA CELL, AND NORMAL OVARIAN FOLLICULAR APPARATUS. Am J Obstet Gynecol. 1965;92:958–69.

    Article  CAS  PubMed  Google Scholar 

  72. Cheng J, Ruan X, Zhou Q, Li Y, Du J, Jin F, et al. Long-time low-temperature transportation of human ovarian tissue before cryopreservation. Reprod Biomed Online. 2021;43(2):172–83.

    Article  PubMed  Google Scholar 

  73. Cacciottola L, Manavella DD, Amorim CA, Donnez J, Dolmans MM. In vivo characterization of metabolic activity and oxidative stress in grafted human ovarian tissue using microdialysis. Fertil Steril. 2018;110(3):534–e443.

    Article  CAS  PubMed  Google Scholar 

  74. Rahimi G, Isachenko E, Sauer H, Isachenko V, Wartenberg M, Hescheler J, et al. Effect of different vitrification protocols for human ovarian tissue on reactive oxygen species and apoptosis. Reprod Fertil Dev. 2003;15(6):343–9.

    Article  PubMed  Google Scholar 

  75. Tavakoli A, Aliakbari F, Soleimani Mehranjani M. Kisspeptin decreases the adverse effects of human ovarian vitrification by regulating ROS-related apoptotic occurrences. Zygote (Cambridge England). 2023;31(6):537–43.

    Article  CAS  PubMed  Google Scholar 

  76. Aubard Y, Piver P, Cogni Y, Fermeaux V, Poulin N, Driancourt MA. Orthotopic and heterotopic autografts of frozen-thawed ovarian cortex in sheep. Hum Reprod. 1999;14(8):2149–54.

    Article  CAS  PubMed  Google Scholar 

  77. Poirot C, Vacher-Lavenu MC, Helardot P, Guibert J, Brugières L, Jouannet P. Human ovarian tissue cryopreservation: indications and feasibility. Hum Reprod. 2002;17(6):1447–52.

    Article  PubMed  Google Scholar 

  78. Schmidt KL, Byskov AG, Nyboe Andersen A, Müller J, Yding Andersen C. Density and distribution of primordial follicles in single pieces of cortex from 21 patients and in individual pieces of cortex from three entire human ovaries. Hum Reprod (Oxford England). 2003;18(6):1158–64.

    Article  CAS  Google Scholar 

  79. Gook DA, Edgar DH, Stern C. Effect of cooling rate and dehydration regimen on the histological appearance of human ovarian cortex following cryopreservation in 1, 2-propanediol. Hum Reprod (Oxford, England). 1999;14(8):2061–8.

  80. Isachenko V, Isachenko E, Woriedh M, Lapidus I, Weiss JM. Vitrification and conventional freezing of human ovarian tissue: follicles formation, hormone production and gene expression. Mol Hum Reprod. 2009;24:i16.

    Google Scholar 

  81. Tao T, Del Valle A. Human oocyte and ovarian tissue cryopreservation and its application. J Assist Reprod Genet. 2008;25(7):287–96.

    Article  PubMed  PubMed Central  Google Scholar 

  82. Rodrigues RB, Uczay M, Brito VB, Godoy AC, Moura DJ, Vogel C, et al. Oxidative stress and DNA damage of zebrafish sperm at different stages of the cryopreservation process. Zebrafish. 2021;18(2):97–109.

    Article  CAS  PubMed  Google Scholar 

  83. Pereira BC, Ortiz I, Dorado J, Consuegra C, Diaz-Jimenez M, Demyda-Peyras S, et al. Evaluation of DNA damage of mare granulosa cells before and after Cryopreservation using a chromatin dispersion test. J Equine Vet Sci. 2019;72:28–30.

    Article  PubMed  Google Scholar 

  84. Chetverikova EP. [DNA damage by active oxygen species in cryopreservation and the antioxydative properties of cryoprotectants]. Biofizika. 2012;57(2):368–76.

    CAS  PubMed  Google Scholar 

  85. Adashi EY. Endocrinology of the ovary. Hum Reprod (Oxford England). 1994;9(5):815–27.

  86. Ouni E, Bouzin C, Dolmans MM, Marbaix E, Pyr Dit Ruys S, Vertommen D, et al. Spatiotemporal changes in mechanical matrisome components of the human ovary from prepuberty to menopause. Hum Reprod (Oxford England). 2020;35(6):1391–410.

    Article  CAS  Google Scholar 

  87. Wang S, Zheng Y, Li J, Yu Y, Zhang W, Song M, et al. Single-Cell Transcriptomic Atlas Primate Ovarian Aging Cell. 2020;180(3):585–e60019.

    CAS  Google Scholar 

  88. Meryman HT. Cryoprotective agents. Cryobiology. 1971;8(2):173–83.

    Article  CAS  PubMed  Google Scholar 

  89. Elliott GD, Wang S, Fuller BJ, Cryoprotectants. A review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures. Cryobiology. 2017;76:74–91.

    Article  CAS  PubMed  Google Scholar 

  90. Hagedorn J, Alkurdi G, Barker SA, Brunotte R, Deeb T, Hubenia O, et al. The technology in cryotechnology. Cryo Lett. 2023;44(6):314–26.

    Article  Google Scholar 

  91. Murray KA, Gibson MI. Chemical approaches to cryopreservation. Nat Rev Chem. 2022;6(8):579–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  92. Awan M, Buriak I, Fleck R, Fuller B, Goltsev A, Kerby J, et al. Dimethyl sulfoxide: a central player since the dawn of cryobiology, is efficacy balanced by toxicity? Regen Med. 2020;15(3):1463–91.

    Article  CAS  PubMed  Google Scholar 

  93. Hovatta O. Methods for cryopreservation of human ovarian tissue. Reprod Biomed Online. 2005;10(6):729–34.

    Article  PubMed  Google Scholar 

  94. Tuten Sevim E, Arat S. Combining dimethyl sulphoxide (DMSO) with different cryoprotectants ensures better cartilage cell cryopreservation. Cryo Lett. 2021;42(4):220–6.

    Google Scholar 

  95. Liu M, Chen C, Liang L, Yu C, Guo B, Zhang H, et al. A biocompatible cell cryoprotectant based on sulfoxide-containing amino acids: mechanism and application. J Mater Chem B. 2023;11(11):2504–17.

    Article  PubMed  Google Scholar 

  96. Whaley D, Damyar K, Witek RP, Mendoza A, Alexander M, Lakey JR. Cryopreservation: an overview of principles and cell-specific considerations. Cell Transpl. 2021;30:963689721999617.

    Article  Google Scholar 

  97. von Bomhard A, Elsässer A, Ritschl LM, Schwarz S, Rotter N. Cryopreservation of endothelial cells in various Cryoprotective agents and media - vitrification versus slow freezing methods. PLoS ONE. 2016;11(2):e0149660.

    Article  Google Scholar 

  98. Oldenhof H, Bigalk J, Hettel C, de Oliveira Barros L, Sydykov B, Bajcsy ÁC, et al. Stallion sperm cryopreservation using various Permeating agents: interplay between concentration and cooling rate. Biopreserv Biobank. 2017;15(5):422–31.

    Article  CAS  PubMed  Google Scholar 

  99. El Cury-Silva T, Nunes MEG, Casalechi M, Comim FV, Rodrigues JK, Reis FM. Cryoprotectant agents for ovarian tissue vitrification: systematic review. Cryobiology. 2021;103:7–14.

    Article  PubMed  Google Scholar 

  100. Hovatta O, Silye R, Krausz T, Abir R, Margara R, Trew G, et al. Cryopreservation of human ovarian tissue using dimethylsulphoxide and propanediol-sucrose as cryoprotectants. Hum Reprod. 1996;11(6):1268–72.

    Article  CAS  PubMed  Google Scholar 

  101. Kasai M, Niwa K, Iritani A. Effects of various cryoprotective agents on the survival of unfrozen and frozen mouse embryos. J Reprod Fertil. 1981;63(1):175–80.

    Article  CAS  PubMed  Google Scholar 

  102. Toishibekov YM, Kazybayeva AS, Assanova YA, Nurkenov TT, Toishybek DY. 44 The effect of various cryoprotective agents and slow cooling rate on viability of goat ovarian tissue. Reprod Fertil Dev. 2021;34(2):257.

Download references

Acknowledgements

We would like to thank all authors who provided published data for our meta-analysis.

Funding

Open Access funding enabled and organized by Projekt DEAL.

These investigations were supported by the China Scholarship Council for Qingduo Kong (No. 202308230130) and Cheng Pei (No. 202208080057).

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Medical Faculty, Cologne University, 50931, Cologne, Germany

    Qingduo Kong, Cheng Pei, Gohar Rahimi, Peter Mallmann & Volodimir Isachenko

  2. Medizinisches Versorgungszentrum AMEDES für IVF- und Pränatalmedizin in Köln GmbH, 50968, Cologne, Germany

    Gohar Rahimi

Authors
  1. Qingduo Kong
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Cheng Pei
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Gohar Rahimi
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Peter Mallmann
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Volodimir Isachenko
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

QDK contributed to the design of the meta-analysis, data analysis, and the completion of the article. CP, GR, and PR contributed to the document selection and the inclusion and exclusion criteria. VI contributed to the design and review of the meta-analysis. All authors approved the final manuscript.

Corresponding author

Correspondence to Volodimir Isachenko.

Ethics declarations

Ethics approval and consent to participate

This study does not contain any studies with human participants or animals performed.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kong, Q., Pei, C., Rahimi, G. et al. Comparison of the quality of ovarian tissue cryopreservation by conventional slow cryopreservation and vitrification—a systematic review and meta-analysis. J Ovarian Res 18, 62 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13048-024-01561-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s13048-024-01561-7

Keywords

Journal of Ovarian Research

ISSN: 1757-2215

Contact us