An Iranian scientist managed to develop a new method to propagate human spermatogonial stem cells from small testicular biopsies to obtain adequate number of cells for successful transplantation for 18000-fold in vitro, leading to protection of the fertility of immature boys suffering from cancer.
The research carried out jointly with Amsterdam University in Holland is the first developed method in the world. (see related research paper)
Hooman Sadri-Ardekani, Ph.D in Reproductive Medicine and a Professor in Avicenna Institute-ACECR told ISNA, "Young boys treated with high-dose chemotherapy are often confronted with infertility once they reach adulthood and cryopreserving testicular tissue before chemotherapy and auto transplantation of spermatogonial stem cells at a later stage could theoretically allow for restoration of fertility."
He stressed project was worked on from April 2007 to July 2009 using testis material donated by 6 adult men who underwent orchiectomy as part of prostate cancer treatment. Testicular cells were isolated and cultured in supplemented StemPro medium, germline stem cell clusters that arose were subcultured on human placental laminin-coated dishes in the same medium.
Sadri-Ardekani continued, "Presence of spermatogonia was determined by reverse transcriptase polymerase chain reaction and immunofluorescence for spermatogonial markers. To test for the presence of functional spermatogonial stem cell in culture, xenotransplantation to tests of immunodeficient mice was performed and migrated human spermatogonial stem cells after transplantation was detected. The numbers of colonized spermatogonial stem cells transplanted at early and later points during culture were counted to determine the propagation."
He referred to the results of the study, noting testicular cells could be cultured and propagated in 15 weeks and germline stem cell clusters arose in testicular cell culture from all 6 men and could be subcultured and propagated in 28 weeks. Expression of spermatogonial markers on both the RNA and protein level was maintained throughout the entire culture period. In 4 of 6 men, Xenotransplantaion to mice demonstrated the presence of functional spermatogonial stem cells, even after prolonged in vitro culture. Spermatogonial stem cell numbers increased by 53-fold within 19 days in the testicular cell culture and increased by 18450-fold within 64 days in the germline stem cell subculture and finally long-term culture and propagation of human spermatogonial stem cells in vitro could be possible.
... Payvand News - 02/05/12 ... --