Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences of Yazd
Abstract
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
