Applying of STO as feeder layer for Rooster Spermatogonial Stem cell culture

The objective of this study was to evaluate the effect of STO feeder layer on prepubertal Rhode Island Red rooster SSCs culture and proliferation in vitro. testis cells, separated cells from prepubertal Rhode Island Red chicken 4-8 weeks, and cultivated with 2 treatment and 3 replications in the presence of bFGF and LIF growth factors on the 4well (2 well were covered with feeder layer) separately. SSCs colonies appear at 5th day of culture. The number of SSCs colonies, cells/colony and colony area was measured in days 7 and 10 in both treatment. In order to evaluate SSCs gene (C-KIT) expression on the 10 day of in both treatment RT-PCR test was used. Data analysis was using by SAS software and the difference between the means was examined by using the general linear model (GLM) procedure and the Duncan test. P < 0. 0 5 was considered significant and. The result of the colony assay at the 7th day revealed a higher colony numbers as well as higher cell number/colony and colony area on STO surface than colonies grown in surface without feeder layer significantly (P≤0.05).in contrast, the result of the colony assay at the 10th were declined in both treatment, According to negative expression for gene, colonies might be composed of SSCs. In conclusion these results indicated that the use of STO feeder layer influenced SSCs proliferation and maintenance of the prepubertal rooster in short-term culture.