In Vitro Spermatogenesis Of Cryopreserved Testicular Tissue For Male Infertility

Aggressive chemotherapy may lead to permanent male infertility. Prepubertal males do not generate sperm, but their testes do contain spermatogonial cells (SPGCs) that could be used for fertility preservation. In the present study, we examined the effect of busulfan (BU) on the SPGCs of immature mice, and the possible induction of the survivor SPGCs to develop spermatogenesis in 3D in-vitro culture. Immature mice were injected with BU, and after 0.5–12 weeks, their testes were weighed and evaluated histologically compared to the control mice. The spermatogonial cells [Sal-like protein 4 (SALL4) and VASA (a member of the DEAD box protein family) in the testicular tissue were counted/seminiferous tubule (ST). The cells from the STs were enzymatically isolated and cultured in vitro. Our results showed a significant decrease in the testicular weight of the BU-treated mice compared to the control. This was in parallel to a significant increase in the number of severely damaged STs, and a decrease in the number of SALL4 and VASA/STs compared to the control. The cultures of the isolated cells from the STs of the BU-treated mice showed a development of colonies and meiotic and post-meiotic cells after four weeks of culture. The addition of homogenates from adult GFP mice to those cultures induced the development of sperm-like cells after four weeks of culture. This is the first study demonstrating the presence of biologically active spermatogonial cells in the testicular tissue of BU-treated immature mice, and their capacity to develop sperm-like cells in vitro.

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In vitro spermatogenesis and fertilization from cryopreserved murine testicular tissue

Nature Reviews Urology ◽

10.1038/nrurol.2014.174 ◽

2014 ◽

Vol 11 (8) ◽

pp. 421-421 ◽

Cited By ~ 1

Author(s):

Robert Phillips

Keyword(s):

Testicular Tissue ◽

In Vitro Spermatogenesis

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In vitro spermatogenesis: The dawn of a new era in the study of male infertility

International Journal of Urology ◽

10.1111/j.1442-2042.2012.02979.x ◽

2012 ◽

Vol 19 (4) ◽

pp. 282-283 ◽

Cited By ~ 3

Author(s):

Takehiko Ogawa

Keyword(s):

Male Infertility ◽

New Era ◽

In Vitro Spermatogenesis

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A controlled study between the use of gamete intrafallopian transfer (GIFT) and in vitro fertilization and embryo transfer in the management of idiopathic and male infertility

Fertility and Sterility ◽

10.1016/s0015-0282(16)59471-0 ◽

1987 ◽

Vol 48 (4) ◽

pp. 605-607 ◽

Cited By ~ 43

Author(s):

John Leeton ◽

Peter Rogers ◽

Catriona Caro ◽

David Healy ◽

Chris Yates

Keyword(s):

In Vitro Fertilization ◽

Male Infertility ◽

Embryo Transfer ◽

Controlled Study ◽

Gamete Intrafallopian Transfer ◽

Vitro Fertilization

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Cellular Therapy via Spermatogonial Stem Cells for Treating Impaired Spermatogenesis, Non-Obstructive Azoospermia

Cells ◽

10.3390/cells10071779 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1779

Author(s):

Nesma E. Abdelaal ◽

Bereket Molla Tanga ◽

Mai Abdelgawad ◽

Sahar Allam ◽

Mostafa Fathi ◽

...

Keyword(s):

Stem Cells ◽

Male Infertility ◽

Cellular Therapy ◽

Antihypertensive Drugs ◽

Testicular Biopsy ◽

Testicular Tissue ◽

Spermatogonial Stem Cells ◽

Obstructive Azoospermia ◽

Promising Alternative ◽

Major Health Problem

Male infertility is a major health problem affecting about 8–12% of couples worldwide. Spermatogenesis starts in the early fetus and completes after puberty, passing through different stages. Male infertility can result from primary or congenital, acquired, or idiopathic causes. The absence of sperm in semen, or azoospermia, results from non-obstructive causes (pretesticular and testicular), and post-testicular obstructive causes. Several medications such as antihypertensive drugs, antidepressants, chemotherapy, and radiotherapy could lead to impaired spermatogenesis and lead to a non-obstructive azoospermia. Spermatogonial stem cells (SSCs) are the basis for spermatogenesis and fertility in men. SSCs are characterized by their capacity to maintain the self-renewal process and differentiation into spermatozoa throughout the male reproductive life and transmit genetic information to the next generation. SSCs originate from gonocytes in the postnatal testis, which originate from long-lived primordial germ cells during embryonic development. The treatment of infertility in males has a poor prognosis. However, SSCs are viewed as a promising alternative for the regeneration of the impaired or damaged spermatogenesis. SSC transplantation is a promising technique for male infertility treatment and restoration of spermatogenesis in the case of degenerative diseases such as cancer, radiotherapy, and chemotherapy. The process involves isolation of SSCs and cryopreservation from a testicular biopsy before starting cancer treatment, followed by intra-testicular stem cell transplantation. In general, treatment for male infertility, even with SSC transplantation, still has several obstacles. The efficiency of cryopreservation, exclusion of malignant cells contamination in cancer patients, and socio-cultural attitudes remain major challenges to the wider application of SSCs as alternatives. Furthermore, there are limitations in experience and knowledge regarding cryopreservation of SSCs. However, the level of infrastructure or availability of regulatory approval to process and preserve testicular tissue makes them tangible and accurate therapy options for male infertility caused by non-obstructive azoospermia, though in their infancy, at least to date.

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The Expression Levels and Cellular Localization of Pigment Epithelium Derived Factor (PEDF) in Mouse Testis: Its Possible Involvement in the Differentiation of Spermatogonial Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22031147 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1147

Author(s):

Noy Bagdadi ◽

Alaa Sawaied ◽

Ali AbuMadighem ◽

Eitan Lunenfeld ◽

Mahmoud Huleihel

Keyword(s):

Cellular Localization ◽

Pigment Epithelium ◽

Testicular Tissue ◽

Seminiferous Tubules ◽

Target Cells ◽

Isolated Cells ◽

Expression Levels ◽

Cellular Origin ◽

Pigment Epithelium Derived Factor

Pigment epithelium derived factor (PEDF) is a multifunctional secretory soluble glycoprotein that belongs to the serine protease inhibitor (serpin) family. It was reported to have neurotrophic, anti-angiogenic and anti-tumorigenic activity. Recently, PEDF was found in testicular peritubular cells and it was assumed to be involved in the avascular nature of seminiferous tubules. The aim of this study was to determine the cellular origin, expression levels and target cells of PEDF in testicular tissue of immature and adult mice under physiological conditions, and to explore its possible role in the process of spermatogenesis in vitro. Using immunofluorescence staining, we showed that PEDF was localized in spermatogenic cells at different stages of development as well as in the somatic cells of the testis. Its protein levels in testicular homogenates and Sertoli cells supernatant showed a significant decrease with age. PEDF receptor (PEDF-R) was localized within the seminiferous tubule cells and in the interstitial cells compartment. Its RNA expression levels showed an increase with age until 8 weeks followed by a decrease. RNA levels of PEDF-R showed the opposite trend of the protein. Addition of PEDF to cultures of isolated cells from the seminiferous tubules did not changed their proliferation rate, however, a significant increase was observed in number of meiotic/post meiotic cells at 1000 ng/mL of PEDF; indicating an in vitro differentiation effect. This study may suggest a role for PEDF in the process of spermatogenesis.

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