Cell Biology: Research & TherapyISSN: 2324-9293

All submissions of the EM system will be redirected to Online Manuscript Submission System. Authors are requested to submit articles directly to Online Manuscript Submission System of respective journal.

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:
[email protected]

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.

Keywords: Alpha modified minimum essential medium; Dulbecco’s modified eagle’s medium; Mesenchymal stem cells; Flow cytometry

Track Your Manuscript

Share This Page

Associations