Optimization of physiological xenofree molecularly defined media and matrices to maintain human embryonic stem cell pluripotency.
We describe in this chapter the development of a xenofree molecularly defined medium, SBX, associated with xenofree matrices, to maintain human embryonic stem cell (hESC) pluripotency as determined by phenotypic, functional and TLDA studies. This simple, inexpensive, and more physiological culture condition has been chosen because (1) it is xenofree and molecularly defined; it is devoid of albumin, which is a carrier of undefined molecules; (2) it maintains pluripotency, but very significantly reduces differentiation gene expression during hESC self-renewal, as compared to the widely used culture conditions tested so far; and (3) it can be further improved by replacing high concentrations of expensive additives by physiological concentrations of new factors. Xenofree molecularly defined media and matrices represent valuable tools for elucidating still unknown functions of numerous embryonic genes using more physiological culture conditions. These genes encode potential new factors controlling hESC self-renewal and pluripotency.
Peiffer
Isabelle
I
Centre National de la Recherche Scientifique, Institut André Lwoff, Villejuif, France.
Barbet
Romain
R
Hatzfeld
Antoinette
A
Li
Ma-Lin
ML
Hatzfeld
Jacques A
JA
D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000235:genetics; D003470:Culture Media / Q000737:chemistry; D016895:Culture Media, Serum-Free / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism
D015415:Biomarkers; D003470:Culture Media; D016895:Culture Media, Serum-Free