Isolation and Enrichment of Spermatogonial Stem Cells with MACS in Cattle Calves by Autopsy in Vitro

Abstract

Spermatognial stem cells (SSCs) are unipotent adult stem cells which form the foundation of spermatogenesis for continuous production of sperms throughout the animal life. Due to their rare availability in the testis, we decided to target CD9 surface marker and magneticactivated cells sorting MACS for enrichment of testicular cell suspension with the cells of stem cell properties. We have successfully characterized the isolated and enriched SSCs population with the Fluorescein isothiocyanate FITC tagged secondary antibody under fluorescent microscope at 500 nm wavelength. Isolated Cells could form the colonies resembling to SSCs colonies after one week of culturing in DMEM along with 10% ABS at 37 0 C temperature, 5% CO 2 and 90%NO 2 . α-N-acetylgalactosamine (GalNAc) is reported to be present in SSCs. We have used FITC-DBA lectin to characterize the SSCs colonies after 14 days of culture. Our findings suggest the use of CD9 surface marker and MACS technology for the enrichment of SSCs and DBA for characterizing SSC colonies.All copyrights reserved to Nexus® academic publishers