Deciphering the importance of three key media components in human embryonic stem cell cultures

• Published in: Biotechnology letters Vol. 28; no. 7; pp. 491 - 495
• Main Authors: DING, Vanessa, CHOO, Andre B. H, OH, Steve K. W
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.04.2006; Springer Nature B.V
• Subjects: Biological and medical sciences; Biotechnology; Cell Culture Techniques - methods; Cell Proliferation - drug effects; Cell Survival - drug effects; Culture Media - chemistry; Culture Media - metabolism; Culture Media, Serum-Free - metabolism; Fibroblast Growth Factor 2 - chemistry; Fibroblast Growth Factor 2 - pharmacology; Fundamental and applied biological sciences. Psychology; Humans; Stem cells; Stem Cells - cytology; Stem Cells - drug effects; Stem Cells - physiology; Tissue Engineering - methods; Human; Cell culture; Serum free medium; stem cells; Embryo; serum-free media; Stem cell; feeder-free culture; human embryonic stem cells
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-006-0005-8

Development of a serum free, feeder-free (SFFF) culture platform for human embryonic stem cells (hESC) will be important for the expansion of hESC for future cell therapy applications. However, currently, culture of hESC consists of a combination of basal media, basic fibroblast growth factor (bFGF), serum replacer (SR) and conditioned media (CM) from feeders, and it is unclear which components of the mixture are absolutely critical in the maintenance of hESC. To evaluate the relative contributions of these media components in the development of SFFF culture, each was systematically eliminated and pluripotency assayed by dual embryonic stem cell markers, Oct-4 and TRA-1-60. We concluded that SR was the most critical component in the platform, followed by bFGF and CM produced by feeders, where down-regulation of Oct-4 occurred after 2, 5 and 5 passages, respectively, upon their withdrawal from the complete media.