[bFGF-stimulated MEF-conditioned medium is capable of maintaining human embryonic stem cells].
4 ng/ml bFGF is indispensable for hESC cultured on mouse embryonic fibroblasts (MEF), withdrawal of bFGF drives the hESC to differentiate. In order to exploit effect of bFGF on MEF, we collected a series of MEF conditioned medium (bFGF-MCM) by co-culturing MEF with increasing bFGF concentrations: 0.03, 0.1, 0.3, 1 and 4 ng/ml. The primitivity of hESC cultured in bFGF-MCM was estimated by morphology and alkaline phosphatase staining. Compared with the control medium (medium conditioned without bFGF: MCM), percentage of undifferentiated colony was increased from 23% to 29%, 44%, 74%, 77% and 78%, respectively. However, percentage of undifferentiated colony in the blank medium (medium conditioned with bFGF but without MEF: bFGF-SR) was from 13% to 31%. This indicated that low concentration of bFGF acted on MEF and stimulated MEF producing effective conditioned medium for maintaining hESC. To identify active elements in the effective conditioned medium can help to understand mechanisms of hESC self-renewal.
Zhou
Yi Ping
YP
Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming 650031, China.
Rochat
Anne
A
Hatzfeld
Antoinette
A
Peiffer
Isabelle
I
Barbet
Romain
R
Hatzfeld
Jacques
J
Li
Ma Lin
ML
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D013997:Time Factors
D017077:Culture Media, Conditioned; D016222:Fibroblast Growth Factor 2