Does prepubertal testicular tissue vitrification influence spermatogonial stem cells (SSCs) viability?

Gholami, Mohammadreza; Hemadi, Masoud; Saki, Ghasem; Zendedel, Abolfazl; Khodadadi, Ali; Mohammadi-Asl, Javad

doi:10.1007/s10815-013-0050-x

Cited by 20 publications

(14 citation statements)

References 45 publications

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“…(2012), Dumont et al . (2015)2.1 M DMSO + 2.7 M EG + 20% FBS + 0.5 M sucroseVial, LN2Preservation of SSCsGholami et al . (2013)Stem Cell KeepVial, LN2Offspring using spz from organotypic cultureYokonishi et al .…”

Section: Resultsmentioning

confidence: 99%

“…Some of these present interesting and promising results on post-thaw cell and tissue integrity, and show the ability to purify viable SSCs for further use (Milazzo et al ., 2008; Gouk et al ., 2011; Wu et al ., 2011, 2014; Unni et al ., 2012; Gholami et al ., 2013; Pacchiarotti et al ., 2013; Yango et al ., 2014; Zhang et al ., 2015). However, these evaluations are merely indicative for the quality of the cryopreservation protocol.…”

Section: Resultsmentioning

confidence: 99%

“…However, two closed vitrification systems have been described for TT cryopreservation with exciting results. Firstly, vitrification of immature mouse TT was successfully performed by simply plunging the vial containing the CPA-submerged samples in LN 2 (Gholami et al ., 2013; Yokonishi et al ., 2014). Secondly, vitrification was achieved by solid-surface vitrification.…”

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

167

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%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

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

Onofre

Baert

Faes

et al. 2016

Hum. Reprod. Update

167

116

View full text Add to dashboard Cite

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“…All Pubmed and Embase articles depicting cryopreservation and gene expression analysis of living human and animal cells and tissue were assessed and revealed six genes of which the expression is known to respond to cryopreservation conditions: two heat shock protein genes ( HSPA1A [16], HSP27 [17]) and four apoptosis-related genes ( CASP3 [18], BAX [19], CD95 [20] and MCL1[21] ). The expression of above mentioned genes was either increased or decreased ( HSPA1A , MCL1 ) in comparison to non frozen cells or tissue.…”

Section: Methodsmentioning

confidence: 99%

Functional Tissue Analysis Reveals Successful Cryopreservation of Human Osteoarthritic Synovium

et al. 2016

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Osteoarthritis (OA) is a degenerative joint disease affecting cartilage and is the most common form of arthritis worldwide. One third of OA patients have severe synovitis and less than 10% have no evidence of synovitis. Moreover, synovitis is predictive for more severe disease progression. This offers a target for therapy but more research on the pathophysiological processes in the synovial tissue of these patients is needed. Functional studies performed with synovial tissue will be more approachable when this material, that becomes available by joint replacement surgery, can be stored for later use. We set out to determine the consequences of slow-freezing of human OA synovial tissue. Therefore, we validated a method that can be applied in every routine laboratory and performed a comparative study of five cryoprotective agent (CPA) solutions. To determine possible deleterious cryopreservation-thaw effects on viability, the synovial tissue architecture, metabolic activity, RNA quality, expression of cryopreservation associated stress genes, and expression of OA characteristic disease genes was studied. Furthermore, the biological activity of the cryopreserved tissue was determined by measuring cytokine secretion induced by the TLR ligands lipopolysaccharides and Pam3Cys. Compared to non frozen synovium, no difference in cell and tissue morphology could be identified in the conditions using the CS10, standard and CryoSFM CPA solution for cryopreservation. However, we observed significantly lower preservation of tissue morphology with the Biofreeze and CS2 media. The other viability assays showed trends in the same direction but were not sensitive enough to detect significant differences between conditions. In all assays tested a clearly lower viability was detected in the condition in which synovium was frozen without CPA solution. This detailed analysis showed that OA synovial tissue explants can be cryopreserved while maintaining the morphology, viability and phenotypical response after thawing, offering enhanced opportunities for human in vitro studies.

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“…Several studies utilised immunohistological evaluation to understand the change in expression of proteins specific to germ cells (Kvist et al 2006;Keros et al 2007;Unni et al 2012;Baert et al 2013), Sertoli and peritubular-stromal cells (Keros et al 2007) and cell proliferation (Milazzo et al 2008;Milazzo et al 2010;Unni et al 2012;Baert et al 2013) in cryopreserved testis. A few studies have also utilised changes in RNA expression to understand the effects of cryopreservation on testis tissues (Unni et al 2012;Gholami et al 2013). However, no report has assessed quantitative changes in protein expression in cryopreserved testicular tissues.…”

Section: Introductionmentioning

confidence: 99%

Comparative efficacies of six different media for cryopreservation of immature buffalo (Bubalus bubalis) calf testis

Devi

Makala

Pothana

et al. 2016

Reprod. Fertil. Dev.

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Buffalo calves have a high mortality rate (~80%) in commercial dairies and testis cryopreservation can provide a feasible option for the preservation of germplasm from immature males that die before attaining sexual maturity. The aim of the present study was to evaluate combinations of 10 or 20% dimethylsulfoxide (DMSO) with 0, 20 or 80% fetal bovine serum (FBS) for cryopreservation of immature buffalo testicular tissues, subjected to uncontrolled slow freezing. Tissues cryopreserved in 20% DMSO with 20% FBS (D20S20) showed total, tubular and interstitial cell viability, number of early apoptotic and DNA-damaged cells, surviving germ and proliferating cells and expression of testicular cell-specific proteins (POU class 5 homeobox (POU5F1), vimentin (VIM) and actin α2 (ACTA2)) similar to that of fresh cultured control (FCC; P>0.05). Expression of cytochrome P450, family 11, subfamily A (CYP11A1) protein and testosterone assay showed that only tissues cryopreserved in D20S20 had Leydig cells and secretory functions identical to that of FCC (P>0.05). High expression of superoxide dismutase2 (SOD2), cold-inducible RNA-binding protein (CIRBP) and RNA-binding motif protein3 (RBM3) proteins in cryopreserved tissues indicated involvement of cell signalling pathways regulating cellular protective mechanisms. Similarity in expression of pro-apoptosis proteins transcription factor tumour protein P53 (TP53) and BCL2-associated X protein (BAX) in D20S20 cryopreserved tissues to that of FCC (P>0.05) suggested lower apoptosis and DNA damage as key reasons for superior cryopreservation.

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Does prepubertal testicular tissue vitrification influence spermatogonial stem cells (SSCs) viability?

Abstract: The study indicates that vitrification of prepubertal testicular tissue does not increase the expression profile of apoptosis-related genes such as Bax and Fas in the testicular SSCs consistent with diminished cell apoptotic/necrotic responses and no increasing intracellular LDH leakage.

Cited by 20 publications

References 45 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

Functional Tissue Analysis Reveals Successful Cryopreservation of Human Osteoarthritic Synovium

Comparative efficacies of six different media for cryopreservation of immature buffalo (Bubalus bubalis) calf testis

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