Differentiation of human oligodendrocytes from pluripotent stem cells

作者：Hu Bao Yang; Du Zhong Wei; Zhang Su Chun^*

来源：Nature Protocols, 2009, 4(11): 1614-1622.

DOI：10.1038/nprot.2009.186

We have developed a four-part protocol to differentiate human embryonic stem cells (hESCESCESCs) to oligodendrocyte progenitor cells (OPCOPCOPCs) according to developmental principles. In the first 2 weeks, hESCESCESCs are induced to differentiate into neuroepithelial cells, which form neural tube-like rosettes. In the following 10 d, these neuroepithelial cells are specified to OLOLIG2-expressing progenitors in the presence of retinoic acid (RARA) and sonic hedgehog (SHH). Upon treatment with fibroblast growth factor 2 (FGF2) for another 10 d, these progenitors convert to OLOLIG2 and NKX2.2-expressing pre-OPCOPCOPCs. Finally, the pre-OPCOPCOPCs take 8-9 weeks to differentiate into OPCOPCOPCs, which express additional markers of oligodendrocytes, such as SOSOX10, platelet-derived growth factor receptor alpha (PDGFR alpha) and NG2. The unique aspects of the protocol are the use of FGF2 to promote the differentiation of gliogenic pre-OPCOPCOPCs in the third part and the removal of FGF2 during the transition of pre-OPCOPCOPCs to OPCOPCOPCs. This 3-month differentiation protocol consistently yields OPCOPCOPCs of high purity capable of producing myelin sheaths in vivo.

• 出版日期2009