Enrichment of Rat Spermatogonial Stem Cells Based on BSA gradient method

Vivek Pandey; Ravi Bhushan; Pawan K. Dubey

Volume : VII, Issue : III, March - 2018

Enrichment of Rat Spermatogonial Stem Cells Based on BSA gradient method

Vivek Pandey, Ravi Bhushan, Pawan K. Dubey

Abstract :

Introduction:–

Spermatogonial stem cells (SSCs) consists a small population of germ cells that have self–renewal and differentiation potential may useful in area of regenerative medicine. However, there are no reliable and established procedures to enrich SSCs population from rat testes. Therefore, it is necessary to develop a method for SSC enrichment.

Aim & Objectives:–

To study the potential of BSA gradient for enrichment of SSCs population from rat testes.

Materials and Methods:–

Testes were isolated from 5 to 6–wk–old mature rats of Charles Foster (CF) stain. The seminiferous tubules were digested using two step enzymatic (collagenase Type IV; trypsin; DNase I, and hyaluronidase) digestion method. The contaminating somatic cells (myoid and sertoli cells) were eliminated by filtration of cell lysate. Finally, enrichment of SSCs was done by using different gradient solution of bovine serum albumin (BSA fraction V). The isolated SSCs were characterized by stem cell markers.

Results:–

The 6% BSA solution showed maximum efficiency for enrichment of SSCs as compared to other concentration of BSA. Isolated SSCs were found positive for stem cell markers THY–1 and alkaline phosphatase strengthens the idea that these cells are stem cells.

Conclusion:–

Development of a new technique for SSC isolation and enrichment from wild type testes using 6% BSA solution will facilitate to get maximum SSCs population for therapeutic and research purposes.