Research Article, Cell Biol Henderson Nv Vol: 7 Issue: 1
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 Patel1, Aruna V Vanikar1,2*, Kunal S Gupte1, Jignesh V Patel2 and Lovelesh A Nigam1
1Department of Cell therapy and Regenerative Medicine, Gujarat, India
2Department of Pathology, Laboratory Medicine, Transfusion Services and Immunohematology, Gujarat, India
*Corresponding Author : Aruna V. Vanikar, MD, PhD, FICP, Prof. and HOD
Department of Cell therapy and Regenerative Medicine, G.R. Doshi and K.M. Mehta Institute of Kidney Diseases &Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS)
Civil Hospital Campus, Asarwa, Ahmedabad – 380016, Gujarat, India
Tel: +91 79 22687043/44
Fax: +91 79 2268 5454
E-mail: vanikararuna@yahoo.com
Received: March 21, 2018 Accepted: May 09, 2018 Published: May 14, 2018
Citation: Patel CN, Vanikar AV, Gupte KS, Patel JV, Nigam LA (2018) 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. Cell Biol (Henderson, NV) 7:1. doi: 10.4172/2324-9293.1000140
Abstract
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.
