Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP۹.۵ markers

چکیده مقاله

Introduction Presently the techniques for making transgenic
animals are cumbersome, required costly instruments and
trained man-power. The ability of spermatogonial stem cells
(SSCs) to integrate foreign genes has provided the opportunity
for developing alternate methods for generation of
transgenic animals. One of the big challenges in this field
is development of the methods to identify and purify donor
SSCs by antibody mediated cell sorting.
Purpose The present study was aimed to identify goat subpopulations
of SSCs using polyclonal antibodies against
PGP9.5 and c-kit molecular markers as well as the growth
characteristics of SSCs during short term culture.
Methods One month old goats’ testicular samples were subjected
for immunohistochemical and immunocytochemical
evaluations. The enzymatically isolated SSCs were cultured
in DMEM plus FCS supplemented with (treatment) or without
(control) growth factors (GDNF, LIF, FGF, and EGF) for
2 weeks. At the end of culture the morphological characteristics
of SSCs colonies and immunocytochemical staining
were evaluated.
Results The number and size of colonies in treatment groups
were significantly (P<0.01) higher than corresponding values