摘要
目的:观察体外培养少突胶质前体细胞的增殖和分化规律,探索其获得纯度较高细胞的实验方法。方法:实验于2004-03-01/08-01在军事医学科学院基础医学所实验室完成。取新生SD大鼠脑皮质体外培养,利用改良的振荡分离纯化法,振荡可将长于星形胶质细胞层表面的少突胶质前体细胞分离出来,差速贴壁以去除其他细胞,传代细胞分别用无血清和有血清的培养液培养,免疫组织化学法测表面抗原进行细胞鉴定。结果:可获得纯度大于95%不同发育阶段的少突胶质细胞系细胞或2型星形胶质细胞。少突胶质前体细胞祖细胞A2B5,O4阳性,不成熟少突胶质细胞O4、O1阳性,成熟少突胶质细胞髓鞘碱性蛋白阳性,2型星形胶质细胞胶质纤维酸性蛋白阳性。结论:改良的振荡分离纯化法是获取高纯度少突胶质细胞系细胞的有效方法。
AIM:To observe the regularity of the proliferation and differentiation of oligodendrocyte precursor cells in vitro,and study the experimental methods to obtain high purity oligodendrocyte precursors cells.METHODS:The experiment was finished in the Institute of Basic Medicine,Chinese PLA Academy of Military Medicine from March 1st to August 1st 2004.Cortex of new neonatal SD rat was obtained and cultured in vitro.Oligodendrocyte precursors,which grew on the surface of astrocytes monolayer,were separated by improved agitation method,and were further purified by differential adheion.Oligodendrocyte precursor cells were cultured in serum free medium and serum medium, respectively. Oligodendrocyte lineage cells were identified by different cell surface markers with immunohistochemistry. RESULTS:The oligodendrocyte lineage cells with >95% purity in different developmental stages or type 2 astrocyte were obtained.Oligodendrocyte progenitor cells were identified by A2B5 and O4 antibodies,immature oligodendrocyte, mature oligodendrocyte and type 2 astrocyte were identified by O4 and O1 antibodies,major basic protein. CONCLUSION:Improved separation and purification through agitation is suitable and effective to obtain oligodendrocyte lineage cells.
出处
《中国临床康复》
CSCD
北大核心
2005年第17期36-37,i003,共3页
Chinese Journal of Clinical Rehabilitation
