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大鼠少突胶质前体细胞纯化及原代培养的新方法
李世平,朱将虎,屈艺,郑臻,母得志
0
(四川大学华西第二医院儿科·出生缺陷与相关妇儿疾病教育部重点实验室)
摘要:
【摘要】目的 建立简单、高效的少突胶质前体细胞(OPCs)分离培养方法。方法 取出生2天的SD大鼠大脑皮层,分离消化皮层细胞为单细胞悬液,采用A2B5免疫磁珠提纯OPCs进行体外培养,倒置显微镜观察细胞生长状况,并在第7天采用免疫荧光检测OPCs特异性标志A2B5和O4;培养7天后换为OPCs分化培养基诱导OPCs分化,分化培养7天后,免疫荧光法检测成熟少突胶质细胞特异性标志骨髓碱性蛋白(MBP)。结果 培养过程中观察发现,95%以上的细胞具有双极或三级突起的典型形态,免疫荧光显示纯化培养的OPCs 95%以上表达A2B5和O4;分化培养后95%以上的细胞为MBP阳性的少突胶质细胞。结论 采用磁珠法可以获得高纯度的OPCs,并且细胞具有分化为成熟少突胶质细胞的能力。
关键词:  OPCs  A2B5免疫磁珠  OPCs分化  原代培养
DOI:
基金项目:国家自然科学基金(81330016,81630038,81270724);四川省科技厅基金(2014SZ0149,2016TD0002)
A novel method for the purification and primary culture of oligodendrocyte precursor cells
LI Shiping,ZHU Jianghu,QU Yi,ZHENG Zhen,MU Dezhi
(Department of Pediatrics, Key Laboratory of Obstetric & Gynecologic and Pediatric Diseases and Birth Defects of Ministry of Education, West China Second Hospital)
Abstract:
【Abstract】Objective To establish a simple and effective method for the purification and primary culture of oligodendrocyte precursor cells (OPCs). Methods OPCs were isolated from neonatal SD rat cortex by antiA2B5 microbeads and cultured in vitro. OPCs were induced by to differentiate into mature oligodendrocytes (OLs). The morphology and biomarkers of OLs were examined through immunofluorescence staining. Results The cultured cells exhibited typical OPCs morphology with bipolar and tripolar bumps, and more than 95% of these cells expressed A2B5. After differentiation, the OPCs progressively differentiated into mature oligodendrocytes which specifically expressed myelin basic protein (MBP) Conclusion High quality and purified OPCs were obtained by antiA2B5 microbeads isolation in vitro, and these OPCs can be differentiated into mature oligodendrocytes.
Key words:  OPCs  Anti A2B5 microbeads  OPCs differentiation  Primary culture

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