Culture and expansion of Adipose derived Mesenchymal stem cells in Ovine

Abstract

Culture and expansion of Adipose derived Mesenchymal stem cells (ADMSCs) in Ovine was carried out in this study. Ovine adipose tissue samples were collected from Chennai Corporation slaughter house under sterile condition in normal saline with five per cent povidone iodine, antibiotic and antimycotic solutions. Collected tissue samples were weighed and digested using collagenase enzyme to isolate adipose tissue derived mesenchymal stem cells (ADMSCs). Cell yield and viability of the cells were calculated by using trypan blue exclusion test. The cells were seeded at the density of one million cells in one T25 culture flask in Dulbecco’s modified Eagle’s medium-high glucose (DMEM-HG). On the day of seeding, the cells showed spherical morphology. Plastic adherence was noticed 24 hrs after seeding. Cell expansion was observed after 3 days. At P0 level, 70% confluency was attained on day 14 and the time taken to reach 70% confluency was reduced to 3-4 days in subsequent passages.