Low oxygen tension affects proliferation and senescence of caprine bone marrow mesenchymal stem cells in in vitro culture condition

Abstract

The culture system of bone marrow mesenchymal stem cells (bmMSCs) in the normoxic environment does not imitate the hypoxic milieu of typical biological conditions, thus hypoxic culture conditions may improve survival, and growth attributes of bmMSCs during in vitro culture. Therefore, the present study was conducted at ICAR-CIRG, Makhdoom during year 2020 with the objective to investigate the changes in biological characteristics of cultured caprine bmMSCs (cbmMSCs) including the cellular senescence, survival, rate of proliferation, immuno-phenotypic characteristics, and gene expression pattern in a normal and hypoxic microenvironment condition. For this, cbmMSCs isolated from bone marrow collected from iliac crest were enriched and grown under either hypoxic (5% O2) or normoxic (20% O2) conditions. Thereafter, the outcome of hypoxic (5% O2) culturing of cbmMSCs on growth characteristics, proliferation, senescence, and expression profile of important stemness-associated (OCT-4) and oxidative stress [glutathione peroxidase (GPx1) and copper-zinc superoxide dismutase (CuZnSOD)] marker genes was evaluated. cbmMSCs cultivated in hypoxic conditions showed higher proliferation and decreased population doubling time and senescence-associated β-GAL expression; however, the immune-phenotypic characteristics of the cells remain unchanged. Furthermore, the culture of cbmMSCs in hypoxia increased the expression of OCT-4, GPx1, and CuZnSOD, compared with the cells grown under normoxia. In conclusion, the culture condition with low O2 level improved the growth characteristics and proliferation of cbmMSCs. These outcomes would provide information to formulate strategies for the collection and efficient in vitro expansion of bmMSCs from goats and other farm animals before their downstream applications.