Cryopreservation of testicular tissue before long-term testicular cell culture does not alter in vitro cell dynamics

Baert, Yoni; Braye, Aude; Struijk, Robert B.; Pelt, Ans M.M. van; Goossens, Ellen

doi:10.1016/j.fertnstert.2015.07.1134

Cited by 53 publications

(39 citation statements)

References 37 publications

(63 reference statements)

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“…2007, (2012)HumanMature1.5 M DMSO + 0.15 M sucrose + 10% HSAInsulated container in −80°C, LN2Maintenance of tissue integrity, preservation of cell dynamics in propagation cultureBaert et al . (2013), (2015)1.28 M DMSO + 25% FBSInsulated container in −80°C, LN2Preservation of SSCsYango et al . (2014)VitrificationMouseImmature2.8 M DMSO + 2.8 M EG + 25% HSAStraw, LN2Maintenance of tissue integrity and activity in organotypic tissue cultureCuraba et al .…”

Section: Resultsmentioning

confidence: 99%

“…Similar results to programmed slow freezing were observed with vitrified-thawed immature human TT (Curaba et al ., 2011a; Poels et al ., 2013). In addition, our group has published an uncontrolled slow freezing protocol able to protect the integrity of adult human TT and maintain cell dynamics in long-term propagation culture, with results similar to fresh controls (Baert et al ., 2013, 2015). However, confirmation with immature TT is still warranted.…”

Section: Resultsmentioning

confidence: 99%

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Cryopreservation of testicular tissue or testicular cell suspensions: a pivotal step in fertility preservation

Onofre

Baert

Faes

et al. 2016

Hum. Reprod. Update

162

116

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BACKGROUNDGerm cell depletion caused by chemical or physical toxicity, disease or genetic predisposition can occur at any age. Although semen cryopreservation is the first reflex for preserving male fertility, this cannot help out prepubertal boys. Yet, these boys do have spermatogonial stem cells (SSCs) that able to produce sperm at the start of puberty, which allows them to safeguard their fertility through testicular tissue (TT) cryopreservation. SSC transplantation (SSCT), TT grafting and recent advances in in vitro spermatogenesis have opened new possibilities to restore fertility in humans. However, these techniques are still at a research stage and their efficiency depends on the amount of SSCs available for fertility restoration. Therefore, maintaining the number of SSCs is a critical step in human fertility preservation. Standardizing a successful cryopreservation method for TT and testicular cell suspensions (TCSs) is most important before any clinical application of fertility restoration could be successful.OBJECTIVE AND RATIONALEThis review gives an overview of existing cryopreservation protocols used in different animal models and humans. Cell recovery, cell viability, tissue integrity and functional assays are taken into account. Additionally, biosafety and current perspectives in male fertility preservation are discussed.SEARCH METHODSAn extensive PubMED and MEDline database search was conducted. Relevant studies linked to the topic were identified by the search terms: cryopreservation, male fertility preservation, (immature)testicular tissue, testicular cell suspension, spermatogonial stem cell, gonadotoxicity, radiotherapy and chemotherapy.OUTCOMESThe feasibility of fertility restoration techniques using frozen-thawed TT and TCS has been proven in animal models. Efficient protocols for cryopreserving human TT exist and are currently applied in the clinic. For TCSs, the highest post-thaw viability reported after vitrification is 55.6 ± 23.8%. Yet, functional proof of fertility restoration in the human is lacking. In addition, few to no data are available on the safety aspects inherent to offspring generation with gametes derived from frozen-thawed TT or TCSs. Moreover, clarification is needed on whether it is better to cryopreserve TT or TCS.WIDER IMPLICATIONSFertility restoration techniques are very promising and expected to be implemented in the clinic in the near future. However, inter-center variability needs to be overcome and the gametes produced for reproduction purposes need to be subjected to safety studies. With the perspective of a future clinical application, there is a dire need to optimize and standardize cryopreservation and safety testing before using frozen-thawed TT of TCSs for fertility restoration.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Cryopreservation of testicular tissue or testicular cell suspensions: a pivotal step in fertility preservation

Onofre

Baert

Faes

et al. 2016

Hum. Reprod. Update

162

116

View full text Add to dashboard Cite

show abstract

“…After puberty, most testes have none or very few degenerated germ cells. Even though spermatogenesis is not possible in an androgen-insensitive environment, as science and technology advance, the possibility of obtaining functional gametes from cultured spermatogonial stem cells acquired from cryopreserved immature testicular tissue is slowly becoming a reality [Hermann et al, 2012;Baert et al, 2015;Guo et al, 2015;Huleihel et al, 2015]. Furthermore, CRISPR/Cas9 mediated genome editing technology shows promising applications in gene therapy, possibly one day preventing or treating genetic diseases as AIS directly via genetic intervention [Huang et al, 2017].…”

Section: Discussionmentioning

confidence: 99%

Androgen Insensitivity Syndrome at Prepuberty: Marked Loss of Spermatogonial Cells at Early Childhood and Presence of Gonocytes up to Puberty

Aliberti¹,

Garrido²,

Marino³

et al. 2017

Sex Dev

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Androgen insensitivity syndrome (AIS) is a hereditary condition in patients with a 46,XY karyotype in which loss-of-function mutations of the androgen receptor (AR) gene are responsible for defects in virilization. The aim of this study was to investigate the consequences of the lack of AR activity on germ cell survival and the degree of testicular development reached by these patients by analyzing gonadal tissue from patients with AIS prior to Sertoli cell maturation at puberty. Twenty-three gonads from 13 patients with AIS were assessed and compared to 18 testes from 17 subjects without endocrine disorders. The study of the gonadal structure using conventional microscopy and the ultrastructural characteristics of remnant germ cells using electron microscopy, combined with the immunohistochemical analysis of specific germ cell markers (MAGE-A4 for premeiotic germ cells and of OCT3/4 for gonocytes), enabled us to carry out a thorough investigation of germ cell life in an androgen-insensitive microenvironment throughout prepuberty until young adulthood. Here, we show that germ cell degeneration starts very early, with a marked decrease in number after only 2 years of life, and we demonstrate the permanence of gonocytes in AIS testis samples until puberty, describing 2 different populations. Additionally, our results provide further evidence for the importance of AR signaling in peritubular myoid cells during prepuberty to maintain Sertoli and spermatogonial cell health and survival.

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“…This remains a major challenge in the preservation of fertile animals with superior genetics, preservation of human reproductive health, as well as in the establishment of cryobanks for wildlife (HOLOCH & WALD, 2011;UNNI et al, 2012;THUWANUT et al, 2013;BAERT et al, 2015).…”

Section: Advances and Perspectives In Testicular Cryopreservationmentioning

confidence: 99%

Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

Lima

Silva

2017

Cienc. Rural

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Cryopreservation of testicular tissue enables the maintenance of reproductive capacity in different animal species, and contributes to the formation of gene banks for endangered species. The spermatogonia present in the testes can be grown in vitro and the sperm obtained can be used in artificial breeding programs. This review aimed to describe the main techniques of testicular cryopreservation, the main cryoprotectants used, as well as the progress made in different animal species thus far. In the last decade, significant progress has been made in obtaining viable and functional germ cells from testicular tissue. However, more research is needed to better establish protocols that can be used in clinical practice with various species.

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Cryopreservation of testicular tissue before long-term testicular cell culture does not alter in vitro cell dynamics

Cited by 53 publications

References 37 publications

Cryopreservation of testicular tissue or testicular cell suspensions: a pivotal step in fertility preservation

Cryopreservation of testicular tissue or testicular cell suspensions: a pivotal step in fertility preservation

Androgen Insensitivity Syndrome at Prepuberty: Marked Loss of Spermatogonial Cells at Early Childhood and Presence of Gonocytes up to Puberty

Cryopreservation of testicular tissue: an alternative to maintain the reproductive capacity in different animal species

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