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OJVRTM
Online Journal of Veterinary Research©
Volume
20(2):126-135, 2016
Effects of CSF1 and GDNF on short-term cultivation of sheep spermatogonial stem cells
Peyman Rahimi-Feyli1, Aliasghar Moghaddam1, Parviz Tajik2, Shiva Shafiei2 , Zahra Nikousefat1,
Mojtaba Goli1, Nafiseh Alighazi2,
Reza Narenji-Sani3
1-Department
of clinical Sciences, Faculty of Veterinary Medicine, Razi University,
Kermanshah, 2- Department of Theriogenology,
Faculty of Veterinary Medicine, University of Tehran, 3- Department of clinical
Sciences, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran
ABSTRACT
Rahimi-Feyli P, Moghaddam A, Tajik
P, Shafiei S, Nikousefat Z, Goli M, Alighazi N, Narenji-Sani R., Effects of CSF1 and GDNF on short-term
cultivation of sheep spermatogonialstem cells, Onl J Vet Res., 20(2):126-135,
2016. Spermatogonial stem cells (SSCs) from mammalian testes could be used to
generate transgenic animals and preserve genetic material. We
describe In vitro colony formation 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 by differential plating. Isolated cells were treated with 40 ng/ml
GDNF; 40ng/ml CSF1 and 40 ng/ml GDNF; or
40ng/ml CSF1. Controls received no treatment. Colony forming assay was done at
day 4, 7 and 10 of cultivation. Cell identification was confirmed by
immunocytochemistry against PGP9.5 and light and transmission electron
microscopy (TEM). Number of colonies were greater with GDNF+CSF1 compared with
CSF1 or controls at day 10 (P<0.05). Surface area of colonies in GDNF + CSF1
group were greater than controls on day 4 and 7 and greater than in
CSF1 on day 7 (P<0.05). The cells also expressed PGP9.5. The findings
suggest that simultaneousaddition of 40ng/ml GDNF and CSF1 to culture media may
increase In vitro colony formation of prepubertal sheep
SSCs.
Key
words: Spermatogonial stem cell, CSF1,
GDNF, Sheep
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