【作者】 富赛里； 胡建国； 李莹； 尹岚； 金建强； 徐晓明； 陆佩华；

【Author】 FU Sai-Li~1 HU Jian-Guo~1 LY Ying~1 YIN Lan~1 JIN Jian-Qiang~1 XU Xiao-Ming~(1,2) LU Pei-Hua~1,* 1 Department of Neurobiology,Shanghai Second Medical University,Shanghai 200025; 2 Kentrucky Spinal Cord Injury Research Center,Department of Neurological Surgery,University of Louisville,511S.Floyd Street MDR616,Louisville,KY40292,USA

【机构】 上海第二医科大学神经生物学实验室； 美国肯塔基脊髓损伤研究中心美国路易威尔大学神经外科系；

【摘要】 本研究采用神经胶质瘤细胞株B104培养上清(B104CM)和碱性成纤维细胞生长因子(bFGF),将冷冻复苏的大鼠胚胎脊髓神经干细胞(NSCs)定向诱导为少突胶质前体细胞OPCs。形态学和免疫组化的结果显示,诱导后95%以上的细胞具有双极或多极突起的典型OPCs 形态,并表达A2B5和PDGFR-α等OPCs 标志,所有PDFR-α阳性的OPCs 均不表达β-TublinⅢ,仅少量细胞表达GFAP。在B104CM 和bFGF 共存的培养条件下,悬浮培养的OPCs 可大量增殖形成少突胶质细胞球该细胞球,可通过传代继续扩增且扩增的OPCs 仍能维持其特有的形态和自我增殖的特性。撤去bFGF 和B104CM后,OPCs 能进一步分化为成熟的少突胶质细胞(OLs)或型星形胶质细胞。结果表明,由NSCs 诱导产生的OPCs 在形态、增殖和分化格局等方面均与已报导的存在于胚胎脑区的O-2A 前体细胞相类似。该培养系统可为实验性细胞移植的研究提供丰富的细胞来源。更多还原

【Abstract】 We have previously established the culture system of isolation andcultivation of neural stem cells(NSCs)from rat embryonic brain and spinalcord.In this study,Spinal cord-derived NSCs were induced intoOligodendrocyte Precursor Cells(OPCs)by combination of conditioned mediumfrom B104(BI04CM)neuroblastoma cells and basic fibroblast growth factor (bFGF,10ng/ml).The morphology and immunofluorecence staining showed thatamong induced cells,over 95% displayed bipolar or tri-polar morphology,whichrepresents typical OPCs and expressed markers of OPCs such as A2B5 andPDGFR.Only a few cells expressed simultaneously GFAP among PDGFRpositiv(?)OPCs.While all OPCs were negative for-tubulinⅢ.In the presence of both B104 CMand bFGF,OPC proliferated largely and then formed oligosphere.These cellscould be expanded in vitro through multiple passages and maintained themorphology of OPCs as well as potential of self-renewal.Upon withdrawal ofBIO4CM and bFGF,they differentiated into04/GlaC-positive oligodendrocytes(OLs)or GFAP/A2B5-positive typeⅡastrocytes,respectively.Our resultsindicated that NSCs could be induced to generate self-renewing OPCs.Themorphologyproliferation and differentiation patterns of OPCs are similar tothat of 0-2A progenitor cells in vivo.OPCs can be maintained in anundifferentiated state over multiple divisions as long as both B104CM and bFGFare Present in the medium.Using this culture model system,we are able toobtain sufficient numbers of OPCs for experimental cell therapy studies.更多还原