The comparison between conditioned media and serum-free media in human embryonic stem cell culture and differentiation.
Human embryonic stem cells (hESCs) offer an inexhaustible supply of human somatic cell types through their ability to self-renew while retaining pluripotency. As such, hESC-derived cell types are important for applications ranging from in vitro modeling to therapeutic use. However, for their full potential to be realized, both the growth of the undifferentiated cells and their derivatives must be performed in defined culture conditions. Many research groups maintain hESCs using mouse embryonic fibroblasts (MEF) and MEF conditioned medium (CM). The use of murine systems to support hESCs has been imperative in developing hESC technology; however, they suffer from some major limitations including lack of definition, xenobiotic nature, batch-to-batch variation, and labor-intensive production. Therefore, hESC culture definition is essential if hESC lines, and their derivatives are to be quality assured and manufactured to GMP. We have initiated the process of standardizing hESC tissue culture and have employed two serum-free media: mTeSR (MT) and Stem Pro (SP). hESCs were maintained in a pluripotent state, for over 30 passages using MT and SP. Additionally, we present evidence that hESCs maintained in MT and SP generate equivalent levels of human hepatic endoderm as observed with CM. This data suggests that MT and SP are effective replacements for MEF-CM in hESC culture, contributing to the standardization of hESC in vitro models and ultimately their application.
Hannoun
Zara
Z
Medical Research Council Centre for Regenerative Medicine, University of Edinburgh , Edinburgh, EH16 4SB, UK.
Fletcher
Judy
J
Greenhough
Sebastian
S
Medine
Claire
C
Samuel
Kay
K
Sharma
Ruchi
R
Pryde
Anne
A
Black
James R
JR
Ross
James A
JA
Wilmut
Ian
I
Iredale
John P
JP
Hay
David C
DC
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D005821:Genetic Techniques; D006801:Humans; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology; D015262:Xenobiotics / Q000494:pharmacology
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D015262:Xenobiotics; D003577:Cytochrome P-450 Enzyme System