In vitro Generation of Human Adipose Derived Mesenchymal Stromal Cells Using \(\alpha\) –MEM and DMEM

Abstract

The basic media for cell culture are \(\alpha\)-MEM and DMEM. \(\alpha\)-MEM has lesser and rather optimal glucose and amino acid concentration as compared to DMEM. In our current study we compared the individual effect of these two media on quantity, viability and immunophenotyping of AD-MSC. Adipose tissue was procured from anterior abdominal wall of healthy willing donors. It was divided equally into two tissue culture flasks with culture medium, one containing \(\alpha\)-MEM and the other containing DMEM. It was incubated in collagenase-I. Pellet was re-suspended in centrifuge tubes with phosphate buffered saline and divided into 6 equal parts. Each set was incubated separately for in-vitro generation in 6-well plates. Media were replenished on alternate days and no passaging was done. One well from each set was harvested by trypsinization, every 3^rd day. Sterility, quantity, viability and immunophenotyping (CD45^-/90⁺/73⁺) by flow cytometry were checked. We analyzed the data to determine maximum total cell count, viability, CD45^-CD90⁺ and CD45^-CD73⁺ population in \(\alpha\)-MEM and compared it with that in DMEM.