Microcarrier-Based Expansion of Adipose-Derived Mesenchymal Stem Cells in Shake Flasks

by Khandaker Siddiquee, PhD and Ma Sha, PhD
Volume 12, Issue 4 (Winter 2013/2014)

The expansion of stem cells, including mesenchymal stem cells (MSCs), has been successfully demonstrated using microcarrier-based small bioreactors such as spinner flasks. In this study, we explored a simple alternative for microcarrier-based MSC expansion using conventional shake flasks. This method relies on a new type of shaker with built-in CO2 gas control capability, the New Brunswick™ S41i incubator shaker. The expansion of adipose-derived mesenchymal stem cells (AdMSCs) was compared between shake and spinner flasks containing microcarriers. The AdMSCs were seeded at a density of 3 × 103 cells/cm2 in both setups, each containing 0.5 g of plastic microcarriers and 50 mL of stem cell growth medium. The cell culture experiments were conducted over 12 days with samples collected daily for cell growth, biochemistry, and metabolite analysis. The study revealed that AdMSCs cultured under shake flask conditions achieved excellent growth under 12-day batch-culture conditions. Finally, the AdMSCs expanded using the shake flask method retained high quality stem cell characteristics, as indicated by CD44 and CD90 stem cell marker assays, and the ability of these cells to differentiate into either adipocytes or osteocytes...

Siddiquee K, Sha M. Microcarrier-Based Expansion of Adipose-Derived Mesenchymal Stem Cells in Shake Flasks. BioProcess J, 2014; 12(4): 32-38. http://dx.doi.org/10.12665/J124.Sha.

Posted online January 13, 2014.