Vol. 11, Issue 10 (2022)

Author(s):
A Thakre, NE Gade, OP Mishra, A Patyal, N Rawat, MK Gendley, AK Nandanwar and J Singh

Abstract:
Stem cells are having potential applications in regenerative medicine due to their self- renewal and multilineage differentiation capability. In vitro cell culture induces environmental stress and leads to expression of cellular stress markers. In present study, we isolated and characterized caprine amniotic fluid derived mesenchymal stem cells (cAF- MSCs) to investigate the expression of cellular stress marker genes. Third passage cAF- MSCs were successfully differentiated in osteogenic and adipogenic lineages and it was confirmed by Alzarin red and Oil red O staining respectively. Total RNA was isolated from cAF-MSCs and cDNA was synthesized by M-MLV reverse transcriptase PCR to study the stress marker genes expression. Gene specific primers were synthesized and PCR conditions were standardized to study the expression of heat shock proteins 70 (HSP 70), heat shock proteins 90 (HSP 90) and Cu-Zn superoxide dismutase (Cu-Zn SOD) genes. Results revealed that the cAF-MSCs express all the three genes involved in cell stress defence mechanism including chaperone proteins (heat shock proteins HSP 70 and HSP 90) and regulation of oxidative stress (Cu-Zn superoxide dismutase). The expression of cellular stress marker genes in cAF-MSCs confirms that the cells are under environmental stress in in vitro culture conditions.