Hepatocyte growth factor enhances the generation of high-purity oligodendrocytes from human embryonic stem cells.
Generation of homogeneous oligodendrocytes as donor cells is essential for human embryonic stem cell (hESC)-based cell therapy for demylinating diseases. Herein we present a novel method for efficiently obtaining mature oligodendrocytes from hESCs with high purity (79.7+/-6.9%), using hepatocyte growth factor (HGF) and G5 supplement (containing insulin, transferrin, selenite, biotin, hydrocortisone, basic fibroblast growth factor and epidermal growth factor) in a four-step method. We induced hESCs into neural progenitors (NP) with HGF (5ng/ml) and G5 (1x) supplemented medium in an adherent differentiation system. The purified NPs were amplified in suspension as neurospheres for 1 month, and terminal oligodendrocyte differentiation was then induced by G5 supplement withdrawal and HGF treatment (20ng/ml). The cells generated displayed typical morphologies of mature oligodendrocytes and expressed oligodendrocyte markers O4 and myelin basic protein (MBP). Our result revealed that HGF significantly enhanced the proliferation of hESC-derived NPs and promoted the differentiation as well as the maturation of oligodendrocytes from NPs. Further studies suggest that HGF/c-Met signaling pathway might play an important role in oligodendrocyte differentiation in our system. Our studies provide a means for generating the clinically relevant cell type and a platform for deciphering the molecular mechanisms that control oligodendrocyte differentiation.
Hu
Zhixing
Z
Laboratory of Reproduction Development Biology, Kunming Primate Research Center, Chinese Academy of Sciences, Kunming, Yunnan 650223, China. xingzhihu@gmail.com
Li
Tianqing
T
Zhang
Xiuzhen
X
Chen
Yongchang
Y
D001973:Bromodeoxyuridine; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D009474:Neurons / Q000166:cytology*; D009836:Oligodendroglia / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D017228:Hepatocyte Growth Factor; D001973:Bromodeoxyuridine