Effects of CSF1 and GDNF on short-term cultivation ofsheep spermatogonial stem cells

摘要

Spermatogonial stem cells (SSCs) from mammalian testes could beused to generate transgenic animals and preserve genetic material. We describe In vitro colonyformation of type A spermatogonia from prepubertal lambs in the presence of GDNF and CSF1.Testicular cells were isolated from testes by two-step enzymatic digestions purified bydifferential plating. Isolated cells were treated with 40 ng/ml GDNF; 40ng/ml CSF1 and 40 ng/mlGDNF; or 40ng/ml CSF1. Controls received no treatment. Colony forming assay was done at day4, 7 and 10 of cultivation. Cell identification was confirmed by immunocytochemistry againstPGP9.5 and light and transmission electron microscopy (TEM). Number of colonies were greaterwith GDNF+CSF1 compared with CSF1 or controls at day 10 (P<0.05). Surface area of colonies inGDNF + CSF1 group were greater than controls on day 4 and 7 and greater than in CSF1 on day7 (P<0.05). The cells also expressed PGP9.5. The findings suggest that simultaneous addition of40ng/ml GDNF and CSF1 to culture media may increase In vitro colony formation of prepubertalsheep SSCs.