In vitro culture and characterization of spermatogonial stem cells on Sertoli cell feeder layer in goat (Capra hircus).

ABSTRACT:

Purpose

To develop an efficient protocol for isolation, purification and long-term culture of spermatogonial stem cell (SSC) in goat.

Methods

The isolation of SSC was performed by testicular disaggregation by enzymatic digestion using collagenase IV, trypsin and DNase I. Further SSCs were enriched using Percoll density gradient centrifugation. The purity of SSCs was assessed by immunocytochemistry (ICC) using α6 integrin. The SSCs were co-cultured on Sertoli cell feeder layer. The SSC colonies were characterized by studying the expression of SSC specific markers (viz., α6 integrin and PLZF) using ICC. The abundance of mRNAs encoding the markers of SSC (viz., β1 integrin and Oct-4) and Sertoli cells (viz., vimentin) was also assayed using quantitative real-time PCR (qPCR).

SUBMITTER: Pramod RK 

PROVIDER: S-EPMC4130931 | biostudies-literature | 2014 Aug

REPOSITORIES: biostudies-literature