The Effect of Laminin and Gelatin Extracellular Matrix on Short-Term Cultivation of Neonate Mouse Spermatogonial Stem Cells

Purpose: To compare the effect of laminin and gelatin on short-term culture of spermatogonial stem cells (SSCs) from neonatal mouse testes.Materials and Methods: 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, a6-Integrin and b1-Integrin. Moreover, the colonies were harvested and the percentage of a6-Integrin and β1-Integrin positive cells was assessed by flowcytometery in both groups.Results: Immunostaining analysis showed that our culture system contained SSC colonies as they were positive for a6-Integrin and b1-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 a6-Integrin, b1-Integrin detected by flow cytometry in both groups. Conclusion: laminin as extracellular matrix cause to increase the number of neonate spermatogonial colonies and decrease the area of them (P£0.05).