Efficient Generation of Oligodendrocytes and GABAergic Neurons from Embryonic Stem Cells and Their Application to Spinal Cord Injury Models

Dong-Wook Kim

Yonsei University College of Medicine

Axon demyelination can result from various central nervous system (CNS) disorders such as stroke, spinal cord injury (SCI) and multiple sclerosis. The axon demyelination often contributes to improper nerve conduction and lead to deterioration of nerve function. Oligodendrocytes (OD) are myelinating cells in the CNS that form myelin sheaths around axons to support rapid nerve conduction. Human embryonic stem cells (hESC) are pluripotent and thus can serve as a valuable cell source for generation of OD. In this study, we attempted to make a protocol in which OD can be generated from hESC to treat demyelinated axon after SCI. hESC were grown on mitomycin-C treated STO feeder layer in DMEM/F12 containing 20% serum replacement. Seven days after passage, colonies were used to form embryoid bodies (EB). Four days after EB formation, hESC were differentiated into neural progenitors through 8-day selection and 8-day expansion culture. Most of the hESC had differentiated into neural progenitors. We were able to verify this by probing with an anti-nestin antibody. Neural progenitors were then grown in OP medium for ~10 days to generate oligodendrocyte precursor cells (OPC) and then treated with MO medium for 2~3 weeks to induce OD (Alternatively, we also use “neurosphere method’ to make OPC and OD). The presence of OPC was confirmed by immunocyto-chemistry by using an antibody against A2B5. ~50% of the cells had stained positive for A2B5. Mature OD, which were O1-positive, were also efficiently induced from OPC in MO medium. The ability for the co-cultured OPC to remyelinate neurons has been verified by electron microscopy. In addition to induction of OD, generation of GABAergic neurons from ESC and their application to the pain model of the spinal cord injury will also be introduced in this talk.