Volume 5, Issue 4 (7-2007)
IJRM 2007, 5(4): 109-115 |
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1- Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2- Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran , mansoure@modares.ac.ir
3- Department of Genetics, School of Basic Sciences, Tarbiat Modares University, Tehran, Iran
4- Department of Genetics, Institute of Royan, Tehran, Iran
2- Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran , mansoure@modares.ac.ir
3- Department of Genetics, School of Basic Sciences, Tarbiat Modares University, Tehran, Iran
4- Department of Genetics, Institute of Royan, Tehran, Iran
Abstract: (2874 Views)
Background: The basis of spermatogenesis is the spermatogonial stem cells (SSCs). The concentration of SSCs is very small. However, a system that supports the proliferation and maintenance of SSCs in vitro could be used to preserve and expand SSCs numbers as well as increase success in transplantation. It is a new avenue to restore spermatogenesis in azoospermia subjects.
Objective: Proliferation and enhancement of frozen-thawed SSCs numbers during in vitro culture.
Materials and Methods: Both Sertoli and spermatogonial cells were isolated from adult mouse testes. Frozen-thawed spermatogonial cells were cultured in two groups: simple culture (Experimental 1) and co culture with Sertoli cells (Experimental 2). Also, Fresh cells were considered as control groups: simple culture (control1) and co culture with Sertoli cells (control 2).Assay of the spermatogonial-cell-derived colonies was carried out at the end of each week.
Results: Results indicated that the viability rate of the frozen cells after thawing (68.4±10.2%) was influenced by cryopreservation procedure significantly (p ≤0.001). In addition, the number of the colonies and their diameters in the co-culture system with fresh cells (25.1±5.2 and 205.8±50 µm, respectively) were more than other groups and the differences were significant (p<0.001). Number of the colonies and their diameters in experimental 1(9.5±4.3 and 124±35.9 µm, respectively), experimental 2 (15.6±3.5 and 157.6±41.9µm, respectively) groups were better than control 1 group (3.1±2.2 and 87.5±30.6µm, respectively) and the differences were significant (p<0.001).
Conclusion: We demonstrated that co-culture system with Sertoli cells can increase in vitro colony formation of adult fresh and frozen-thawed spermatogonial cells in mouse.
Objective: Proliferation and enhancement of frozen-thawed SSCs numbers during in vitro culture.
Materials and Methods: Both Sertoli and spermatogonial cells were isolated from adult mouse testes. Frozen-thawed spermatogonial cells were cultured in two groups: simple culture (Experimental 1) and co culture with Sertoli cells (Experimental 2). Also, Fresh cells were considered as control groups: simple culture (control1) and co culture with Sertoli cells (control 2).Assay of the spermatogonial-cell-derived colonies was carried out at the end of each week.
Results: Results indicated that the viability rate of the frozen cells after thawing (68.4±10.2%) was influenced by cryopreservation procedure significantly (p ≤0.001). In addition, the number of the colonies and their diameters in the co-culture system with fresh cells (25.1±5.2 and 205.8±50 µm, respectively) were more than other groups and the differences were significant (p<0.001). Number of the colonies and their diameters in experimental 1(9.5±4.3 and 124±35.9 µm, respectively), experimental 2 (15.6±3.5 and 157.6±41.9µm, respectively) groups were better than control 1 group (3.1±2.2 and 87.5±30.6µm, respectively) and the differences were significant (p<0.001).
Conclusion: We demonstrated that co-culture system with Sertoli cells can increase in vitro colony formation of adult fresh and frozen-thawed spermatogonial cells in mouse.
Type of Study: Original Article |
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