Culture of oligodendrocyte precursor cells (NG2(+)/O1(-)) and oligodendrocytes (NG2(-)/O1(+)) from embryonic rat cerebrum.
I have established a novel culture technique of oligodendrocyte precursor cells (OPC, NG2(+)/O1(-)) and oligodendrocytes (OL, NG2(-)/O1(+)) from embryonic day 16 (E16) rat cerebrum distinguished by morphological and immunocytochemical analyses. This novel protocol does not require immunopanning techniques using specific antibodies. The OPC were isolated by two passages every 7 days and culturing them on Petri dishes in different culture medium at each step to eliminate neurons and astrocytes. The yield of pure OPC and OL was relatively large compared with immunopanning techniques. In addition, to examine myelination processes in vitro, the OL from different stages were co-cultured with primary neurons from E17 rat cerebrum on a feeder layer of rat cerebrum astrocytes. This co-culture system resulted in successful formation of myelin in the presence or absence of astrocytes. This culture system was useful for studying OL lineage and initiation of myelination.
Itoh
Kouichi
K
Department of Molecular Biodynamics, The Tokyo Metropolitan Institute of Medical Science, Tokyo Metropolitan Organization for Medical Research, Rm 408, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8613, Japan. itoh@rinshoken.or.jp
D000818:Animals; D001253:Astrocytes / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000276:immunology; D048429:Cell Size / Q000276:immunology; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D003470:Culture Media / Q000494:pharmacology; D005260:Female; D005333:Fetus; D005455:Fluorescent Antibody Technique / Q000295:instrumentation / Q000379:methods; D008854:Microscopy, Electron; D009186:Myelin Sheath / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D009474:Neurons / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D009836:Oligodendroglia / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*; D013687:Telencephalon / Q000196:embryology* / Q000378:metabolism / Q000648:ultrastructure*
D015415:Biomarkers; D003470:Culture Media