Spermatogonial stem cell preservation and transplantation: from research to clinic

Goossens, Ellen; Saen, Dorien Van; Tournaye, Herman

doi:10.1093/humrep/det039

Cited by 133 publications

(88 citation statements)

References 101 publications

(99 reference statements)

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“…A proposed solution for this problem is the restoration of fertility via SSC transplantation. Since many of the patients in this situation are prepubertal (i.e., do not yet produce sperm), a current approach consists on banking testicular tissue or cell suspensions prior to the oncological treatment and submitting the patient to an autologous transplantation of the preserved material if fertility problems arise later in life [115] (for recent advances in the methods for preservation of fertility on young male human oncological patients see the comprehensive review from Goossens et al [116] , 2013). A word of caution is valid regarding safety issues when malignant diseases are managed with transplantation procedures.…”

Section: Fertility Restorationmentioning

confidence: 99%

“…In experiments with rats, only as few as 20 leukemia cancer cells were enough to produce the disease when transplanted into healthy rat testis [117] . It is unknown to what extent this is reproducible in humans, but as an obvious measure, cell suspensions should be screened and positively selected for SSCs and/or negatively selected for cancer cells, through approaches such as FACS or special culture conditions [116] . An alternative approach would be to use testis xenografting or allografting with the hope to restore the spermatogenic process ectopically to a degree just enough to produce a few sperm to be later used with ARTs.…”

Section: Fertility Restorationmentioning

confidence: 99%

See 1 more Smart Citation

Spermatogonial stem cells: Current biotechnological advances in reproduction and regenerative medicine

Aponte

2015

WJSC

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Section: Fertility Restorationmentioning

confidence: 99%

Section: Fertility Restorationmentioning

confidence: 99%

Spermatogonial stem cells: Current biotechnological advances in reproduction and regenerative medicine

Aponte

2015

WJSC

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“…After thawing, SSCs must undergo a maturation process to restore the ability to produce spermatozoa. In vivo transplantation of SSCs or testicular grafting into sterile patients after recovery (Wyns et al, 2010;Goossens et al, 2013), as well as in vitro culture of isolated SSCs or organ culture of testicular fragments (Reuter et al, 2012;Song & Wilkinson, 2012), have been proposed to achieve the complete maturation process of spermatogenesis. In vitro spermatogenesis is particularly interesting to circumvent the re-introduction of malignant cells into patients, such as in cases of leukaemia .…”

Section: Introductionmentioning

confidence: 99%

Assessment of the optimal vitrification protocol for pre-pubertal mice testes leading to successful in vitro production of flagellated spermatozoa

et al. 2015

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SUMMARYTesticular tissue cryopreservation offers the hope of preserved future fertility to pre-pubertal boys with cancer before exposition to gonadotoxic treatments. The objective of this study was to compare controlled slow freezing (CSF) with five vitrification techniques for cryopreservation of murine pre-pubertal testicular tissue and to evaluate the best protocol that could provide a successful completion of spermatogenesis after in vitro maturation. Testicular tissue from 24 mice at 6.5 days post-partum (dpp) was used to compare several vitrification protocols with one another, as well as with a CSF protocol. Toxicity test using additional 12 mice was performed for all cryopreservation solutions. Fresh tissue (FT) from six mice was used as a control. Once the optimal vitrification protocol was selected [the modified solid surface vitrification No. 1 (mSSV 1 )], testes from 18 mice were cultured in vitro for 30 days with (i) fresh, (ii) slow-frozen/thawed and (iii) vitrified/warmed tissues. Testes from six mice at 36.5 dpp were used as controls. At day 30 of in vitro culture, germ cells of the seminiferous tubules showed a high ability to proliferate and elongated spermatids were observed after both freezing techniques, confirming the successful completion of in vitro spermatogenesis. However, after mSSV 1 , the morphological alterations and the percentage of pyknotic seminiferous tubules were lower than CSF (4.67 AE 0.53 vs. 10.1 AE 1.12 and 22.7 AE 2.83% vs. 37.3 AE 4.24% respectively). Moreover, the number of flagellated spermatozoa produced per mg of tissue was higher for mSSV 1 than for CSF (35 AE 3 vs. 9 AE 4 cells), with amounts of secreted testosterone during the culture close to those of FT. The mSSV 1 protocol resulted in success rates better than CSF in maintaining testicular tissue structure, tubular morphology and tissue functions not solely for immediate frozen/thawed tissues but also after a long-term in vitro culture.

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“…The SSCs can serve as a model for better understanding of adult stem cell biology and deciphering the mechanisms that control SSC functions. Recent studies advocate the implication of cryopreservation and transplantation of SSCs in transgenesis [1,3]. However, the exploitation of SSCs is limited due to the inherent problems of isolation, enrichment and establishment of long term culture [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus)

Pramod

Mitra

2014

J Assist Reprod Genet

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Purpose To develop an efficient protocol for isolation, purification and long-term culture of spermatogonial stem cell (SSC) in goat. Methods The isolation of SSC was performed by testicular disaggregation by enzymatic digestion using collagenase IV, trypsin and DNase I. Further SSCs were enriched using Percoll density gradient centrifugation. The purity of SSCs was assessed by immunocytochemistry (ICC) using α6 integrin. The SSCs were co-cultured on Sertoli cell feeder layer. The SSC colonies were characterized by studying the expression of SSC specific markers (viz., α6 integrin and PLZF) using ICC. The abundance of mRNAs encoding the markers of SSC (viz., β1 integrin and Oct-4) and Sertoli cells (viz., vimentin) was also assayed using quantitative real-time PCR (qPCR). Results The viability of isolated testicular cells was>90 % and the Percoll density gradient method resulted in 3.65 folds enrichment with a purity of 82.5 %. Co-culturing of SSCs with Sertoli cell feeder layer allowed the maintenance of stable SSC colonies even after one and half months of culture. The results of ICC analysis showed the expression of α6 integrin and PLZF in almost all the SSC colonies. qPCR analysis revealed a differential expression of mRNAs encoding β1 integrin, Oct-4 and vimentin markers. Conclusion Results of this study demonstrate a simple enzymatic digestion and Percoll density gradient method for isolation and enrichment of SSCs, and suitability of Sertoli cell feeder layer for long term in vitro culture of SSC in goats.Results also suggest the possible application of non-caprine antibodies against SSC specific markers for the identification and subsequent assessment of SSCs in goats.

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Spermatogonial stem cell preservation and transplantation: from research to clinic

Cited by 133 publications

References 101 publications

Spermatogonial stem cells: Current biotechnological advances in reproduction and regenerative medicine

Spermatogonial stem cells: Current biotechnological advances in reproduction and regenerative medicine

Assessment of the optimal vitrification protocol for pre-pubertal mice testes leading to successful in vitro production of flagellated spermatozoa

In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus)

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