Maintaining the male germline: Efficient derivation of ES-like cell colony from spermatogonial cell sources in vitro has been challenging in the treatment of cell therapy Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Morphometric study of human spermatogonial stem cells in receipt mouse testes. To generate ES-like colonies of this experiment, we used co-culture system because; it is assumed that GDNF and other necessary growth factors might be provided by the sertoli cells. This article has been cited by other articles in PMC. Conflict of interest The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

Developmental and Regenerative Potential. Our research emphasized that testicular tissue culture system could support homing of transplanted cells in the testes of recipients. Testicular tissues can successfully support cells to preserve their anatomical and physical structures, because these tissues contain all cell types, including interstitial cells Testicular tissue cut to small pieces after in vitro transplantation and they were placed on agarose gel after transplantation, C. DAPI staining was also done to show all cells. Staining of the cells was verified under a fluorescent microscope, and the cells were washed with PBS.

These cells support the germ cell development by providing structural support, secreting necessary cytokines and growth factors 20 – Spermatogonial stem cells as a source for regenerative medicine. They observed homing of mesenchymal stem cells after 14 days and assumed that an atrophied disc tissue plays chemoattractive activities for mesenchymal stem cells. Materials and methods Testicular cell isolation and culture In this experimental in vitro study protocols for animal care and surgical intervention of neonate mouse in this study were approved by the Institution Animal Care and Use Committee at the University of Baqiyatallah, Tehran, Spermatogonila.


The difference between our study and the much published studies on SCs dedifferentiation to ES-like cells lies in the culture method used 6911 With these valuable features, spermatogonial stem cell research holds great promises in not only the revolution of fertility preservation tbesis, but also in clinical applications. Methods for culturing mouse and rat SSCs in vitro have been established in different laboratories 1631 Tracing DiI two weeks after transplantation in different groups, observing under immunofluorescent microscope.

It reduced consumption of additional growth factors and feeder layer that may cause undesirables genetic changes which might directed to mutation and cancer. Transplantation of spermatogonial stem cells isolated from leukemic spermatotonial restores fertility without inducing leukemia.

Here we report the successful establishment of a simple and cost-effective co- culture system for ES-like cells derivation from neonatal mouse testis. Some male germ cells called spermatogonial stem cells SSCs exist on the basic membrane of seminiferous spermarogonial in spermatognial and participate in spermatogenesis 1.

Welcome at the Research Unit BIOLOGY OF THE TESTIS (BITE) – BITE

Testicular tissue cut to small pieces after in vitro transplantation and they were placed on agarose gel after transplantation, C. On the other hand, testis tissue culture can provide a safe system to induce spermatogenesis out of host body. In addition, the application of stem cells with no ethical problems and minimal intervention in vitro for regenerative medicine will be valuable.

spermatogonial stem cells phd thesis

Autologous spermatogonial stem cell transplantation in man: For example, Potocnik et al. Although the most important role of SSCs is the unlimited production of sperm, they are the unique population of male adult stem cells which have the responsibility of passing genetic material to the next generation 2 – 4. Methods for the quantitative morphologic analysis of tissues.

Open in a separate window. Sertoli cells are the only somatic cells that reside within the seminiferous tubules.



Colonies with dark-red color were considered positive for ALP activity. Therefore, we used co-culture system to supply the critical factors from sertoli cells as well as SCs instead of adding any growth factors or different source of sertoli cells for SCs derived ES-like cells.

Firstly, they detected a homing defect in immature pup recipient testis, demonstrating lack of the blood-testisbarrier BTB. The results indicated that ES-like cell with pluripotency characteristic were generated from freshly isolated spermatogonial cells. Spermatogonial cells were purified by differential plating.

Considering the importance of achieving in vitro spermatogenesis and establishing a tissue culture environment, we aimed to develop a suitable in vitro recipient testis for the homing and resumption of spermatogenesis with human SSCs.

Staining of the cells was verified under a fluorescent microscope, and the cells were washed with PBS. In many studies, researchers have tried to provide natural thessi such as microenvironment and growths factors for cell culture as far as possible.

Dil tracking revealed that majority of the cells in the transplantation groups were Dil positive Fig. Most testis cells were attached to the growing surface two days of postplating.

Results showed expression of associated pluripotency markers, such as Oct4, Nanog and c-Myc, whereas SCs freshly isolated from the testes were negative for these markers except Oct4. Received Dec 20; Accepted Feb 5. Mohammad Hossein AsadiPh. Future of spermatogonial stem cell culture: One of the most important factors that is secreted by sertoli cell is GDNF.

spermatogonial stem cells phd thesis

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