Reza Youssefi - Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
Parviz Tajik - Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran
Mansoureh Movahedin - Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Vahid Akbarinejad - Young Researchers and Elites Club, Roudehen Branch, Islamic Azad University, Roudehen, Iran

Enrichment of cell suspension with germ cells prior to injection into recipient seminiferous tubules is of importance in spermatogonial stem cells (SSCs) transplantation. Knock-out serum replacement (KSR) has been reported to enhance the proliferation of murine SSCs and human embryonic stem cells. The aim of the present study was to investigate the effect of KSR versus fetal bovine serum (FBS) and their interaction on colonization of bovine SSCs in vitro. When FBS (۱۰%) was replaced with KSR (۱۰%), a significant increase in the colonization of SSCs and the expression of Thy۱, as marker for enrichment of SSCs, was observed. It was revealed that the lesser proliferative effect of FBS as well as the greater proliferative impact of KSR on SSCs colonization were not irreversible as cells having been cultured with FBS (۱۰%) for three days with low colonization showed high rate of colonization in response to KSR (۱۰%) and cells having been cultured with KSR (۱۰%) with high colonization experienced low rate of colonization in response to FBS (۱۰%). Further, it was shown that FBS did not contain factors inhibiting SSCs colonization and it simply lacked factors essential for SSCs proliferation because the combination of FBS (۵%) and KSR (۵%) resulted in even greater rate of colonization than did KSR (۱۰%). In conclusion, the present study showed that addition of KSR to culture medium would significantly increase SSCs proliferation.

Bovine, Colonization, Fetal bovine serum, Knock-out serum replacement, Spermatogonial stem cell