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  • 陈美兰,唐中俊,明健,王浩,胡春,蔡季平.大鼠寡突胶质前体细胞的体外培养、鉴定及其生物学特性观察[J].第二军医大学学报,2012,33(2):130-135    [点击复制]
  • CHEN Mei-lan,TANG Zhong-jun,MING Jian,WANG Hao,HU Chun,CAI Ji-ping.Culture, identification and characterization of rat oligodendrocyte precursor cells in vitro[J].Acad J Sec Mil Med Univ,2012,33(2):130-135   [点击复制]
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大鼠寡突胶质前体细胞的体外培养、鉴定及其生物学特性观察
陈美兰,唐中俊,明健,王浩,胡春,蔡季平*
0
(第二军医大学长征医院眼科,上海 200003
*通信作者)
摘要:
目的 探讨体外培养条件下寡突胶质前体细胞增殖、迁移及分化特性。方法 取新生1 d SD大鼠大脑皮质混合胶质细胞培养10 d,分离纯化寡突胶质前体细胞。通过BrdU掺入实验对其增殖能力进行测定,采用经典的Transwell模型和重聚球模型对其迁移能力进行检验,并在分化培养基中观察其分化成为成熟寡突胶质细胞的能力。结果 混合培养胶质细胞在培养10 d后出现明显分层现象。初次振荡后,小胶质细胞脱落显示大量聚集成团的寡突胶质前体细胞散布于星形胶质细胞表面。分离的寡突胶质前体细胞NG2、A2B5表达阳性;阿糖胞苷可抑制其增殖(P<0.01);PDGF可明显促进其迁移(P<0.01);在分化培养条件下,MBP阳性的成熟寡突胶质细胞增多(P<0.01)。结论 寡突胶质前体细胞在体外培养条件下可以保持其增殖、迁移和分化的基本特性。
关键词:  寡突胶质前体细胞  细胞增殖  细胞迁移  细胞分化  细胞培养技术
DOI:10.3724/SP.J.1008.2012.0130
投稿时间:2011-12-06修订日期:2012-02-04
基金项目:
Culture, identification and characterization of rat oligodendrocyte precursor cells in vitro
CHEN Mei-lan,TANG Zhong-jun,MING Jian,WANG Hao,HU Chun,CAI Ji-ping*
(Department of Ophthalmology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
*Corresponding author.)
Abstract:
Objective To observe the proliferation, migration and differentiation of highly enriched oligodendrocyte precursor cells (OPCs) in vitro. Methods OPCs were dissociated and purified by shaking method from cultured glia mixture, which was derived from postnatal 1 day SD rats. BrdU incorporation assay was applied to analyze the proliferation ability of OPCs. Transwell and reaggragration assay were employed to determine the migration ability of OPCs. In differentiation model, the maturation ability of OPCs was examined by immunostaining and Western blotting analysis of myelin basic protein (MBP). Results The stratification of glia mixture occurred 10 days after culture. The microglia was removed by first session of shaking, and there were multiple aggregated OPCs scattered on the surface of astrocyte layer. The separated OPCs were positive for NG2 and A2B5. Ara-C greatly inhibited the proliferation of OPCs as observed by BrdU incorporation assay. PDGF obviously promoted migration of OPCs in both Transwell assay and reaggregation models. The ratio of MBP positive cells (mature oligodendrocytes) was increased in the differentiation medium. Besides, Western blotting analysis showed that MBP in differentiation medium was greatly elevated compared to that in proliferation medium (P<0.01). Conclusion The in vitro cultured and purified OPCs can maintain their fundamental characteristics of proliferation, migration and differentiation.
Key words:  oligodendrocyte precursor cells  cell proliferation  cell migration  cell differentiation  cell culture techniques