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- Title
Comparing the efficiency of different culture systems on proliferation and purification of the spermatogonial stem cells from obstructive azoospermic patients.
- Authors
Zahiri, M.; Movahedin, M.; Mowla, S. J.; Noruzinia, M.
- Abstract
Introduction: Spermatogenesis is a highly organized process that is tightly regulated. Spermatogonial stem cells (SSCs) are responsible for Spermatogenesis. Studying about the biology of SSCs provides better understanding about male infertility, germ cell cancer and male contraception. Materials and Methods: As the mechanisms that involved in human spermatogenesis are complex and unknown, we evaluate different cultural Colonization of isolated human SSCs were studied in various groups during 2 weeks culture. Equal number of cell population in each group was sorted with MACS for GFR-α1 antibody and the other part was not sorted. Both groups were cultured for further one week. Gene specific methylation and quantitative genes expression of pluripotency (Nanog, C-Myc, Oct-4) and specific germ cell) Integrin α6, Integrin β1, PLZF) genes in each stages were evaluated by MSP and quantitative PCR. To revealing functionality, spermatogonial cells from the selected group were transplanted to azoospermia mouse model. Colonization of isolated human SSCs were studied in various groups during 2 weeks culture. Equal number of cell population in each group was sorted with MACS for GFR-α1 antibody and the other part were not sorted. Both groups were cultured for further one week. Gene specific methylation and quantitative genes expression of pluripotency (Nanog, C-Myc, Oct-4) and specific germ cell) Integrin α6, Integrin β 1, PLZF) genes in each stages were evaluated by MSP and quantitative PCR. To revealing functionality, spermatogonial cells from the selected group were transplanted to azoospermia mouse model. Results: The results showed that the number and diameter of colonies in testicular cell suspension was significantly higher than others (p⩽0.05). The expression of germ specific genes in testicular cell suspension and after purification was significantly increased (p⩽0.05). Nanog and C-Myc expression level were significantly decreased in this group (p⩽0.05). There was no significant difference about the expression of Oct-4 among testicular cell suspension and other groups (p>0.05). Conclusion: Gene specific methylation pattern of examined genes didn't show any changes during culture period. Our data from transplantation indicated the homing of the donor derived cells and the presence of human functional sperm. In conclusion our results confirmed that culture of testicular cell suspension and selection of spermatogonial cells could be effective ways for purification and enrichment of the functional human spermatogonial cells.
- Subjects
GERM cells; SPERMATOGENESIS; MALE contraception; CANCER cells; CELL proliferation; BIRTH control
- Publication
Iranian Journal of Reproductive Medicine, 2013, p13
- ISSN
1680-6433
- Publication type
Article