In vitro propagation of spermatogonial stem cells SSCs of Clarias batrachus and production of fertile sperms from the propagating SSCs

Abstract

The in vitro propagation of spermatogonial stem cell is a unique platform to investigate and newlineunderstand the highly complex season specific spermatogenesis process in teleost fishes. There is newlinea knowledge gap about Spermatogonial stem cell biology and information is insufficient to newlineexplore the molecular mechanism behind SSC behavior under in vivo condition. Spermatogonial newlinestem cells (SSC) are adult stem cells capable of transferring genetic information through many newlinegenerations. It is quite impossible to study the spermatogenesis process in vivo. So, in vitro newlinepropagation of SSC is one and the only option to generate knowledge about reproductive biology newlineof teleost fishes. The research about SSCs has been a unique platform to investigate the concept newlineof totipotency. SSCs can produce sperm throughout the lifespan of a male by spermatogenesis newlineprocess and have a very important role in terms of reproduction by inheritance of characters newlinewhich can be independently expressed in off springs. The single cell sperm may be denoted as newlinevector, really conserves the genetic footprint of a species as a sign of evolution and diversity. As newlinecompared to mammals, intense research regarding SSC biology has been carried out in teleostian newlinefishes.The main objective of the present study was to enrich the SSC population from the testis newlineof farmed catfish, Clariasbatrachus (commonly known as magur, walking catfish or air newlinebreathing catfish) and in vitro production of sperm from the propagating spermatogonial cells. newlineThe primary culture of the spermatogonial stem cell was done by treating the testicular cells with newlinecollagenase to obtain mixed population of cells containing spermatids, spermatozoa, blood cellsother somatic cells including motile sperm. To obtain the pure culture of only SSCs, further newlineenrichment was carried out by using Ficoll gradient centrifugation technique followed by newlineMagnetic activated cell sorting (MACS) using Thy1.2 (CD90.2) antibody. The recovery rate of newlinepurified spermatogonial cells counted was about 3×106 newline cells from 6×106