Клеточная биология: исследования и терапия

Comparison of Alpha Modified Eagle’s Minimum Essential Medium with Dulbecco’s Modified Eagle’s Medium For In-Vitro Generation of Human Adipose Tissue Derived Mesenchymal Stem Cells.

Chetan N Patel, Aruna V Vanikar*, Kunal S Gupte, Jignesh V Patel and Lovelesh A Nigam

Background: Alpha modified Eagle’s minimum essential medium (α-MEM) and Dulbecco’s modified Eagle’s medium (DMEM) are the basic media for cell culture. α-MEM has minimum glucose and amino acid concentration than DMEM. We report a study comparing the individual effect of these two media used for generation of adipose tissue derived mesenchymal stem cells (ADMSC), in terms of quantity, viability and immunophenotyping. Material and Methods: Ten grams adipose tissue resected from anterior abdominal wall of 5 willing healthy donors was divided equally into two 75 cm2 tissue culture flasks with 40 ml culture medium, one containing α-MEM and the other containing DMEM. Adipose tissue was minced into tiny pieces, incubated in collagenase-I at 370C for 1 h and centrifuged at 780 rpm for 8 min. Supernatant was removed and pellets collected in 15 ml centrifuge tubes, diluted to 10 ml with phosphate buffered saline and divided into 2 equal parts. Each set was incubated separately in 6-well plates for 15 days for in-vitro generation. Media were replenished on alternate days. Every 3rd day, 1 well from each set was harvested by trypsinization and analysed for sterility, quantity, viability and immunophenotyping (CD45-/90+/73+) by flow cytometry.

Results: Maximum total cell count was 45.66 × 104 cells/ml on thday of culture with 99.6% viability, CD45-CD90+,8.53 × 104 cells/ ml and CD45-CD73+, 5.69 × 104 cells/ml in α-MEM versus 7.5 × 104 cells/ml with 90.5% viability, CD45-CD90+ 1.8 × 104 cells/ml and CD45-CD73+ 1.84×104 cells/ml in DMEM.

Conclusion: α-MEM was superior to DMEM for in-vitro generation of ADMSC.