[ effect of laminin and gelatin extracellular matrix on short-term cultivation of neonate mouse spermatogonial stem cells]

ABSTRACT

To compare the effect of laminin and gelatin on short-term culture of spermatogonial stem cells [SSCs] from neonatal mouse testes. Cell suspension containing SSCs were isolated from testes of 6 day-old mice and cultured in the presence of Glial-derived neuroterophic factor [GDNF], Epidermal Growth Factor [EGF] and Basic Fibroblastic Growth Factor [bFGF] on laminin-and gelatin-coated plates for 9 days. Number and area of colonies were measured in 5th, 7th and 9th days after culturing. At 9th day Immunostaining was used to detect expression of SSC markers, alpha 6-Integrin and beta 1-Integrin. moreover, the colonies were harvested and the percentage of alpha 6-Integrin and beta 1-Integrin positive cells was assessed by flowcytometery in both groups. Immunostaining analysis showed that our culture system contained SSC colonies as they were positive for alpha 6-Integrin and beta 1-Integrin. Additionally, the number of colonies those were formed on laminin were significantly higher in comparison with those of other group. but colony area was higher on gelatin. There was no significant difference in percentage of cells that expressed alpha 6-Integrin, beta 1-Integrin detected by flowcytometry in both groups. laminin as extracellular matrix cause to increase the number of neonate spermatogonial colonies and decrease the area of them [P