改良传代法获取大量高纯度大鼠少突胶质前体细胞  

Modified passage approach for obtaining mass of high purity oligodendrocyte precursor cells of rat

作　　者：陈婧[1] 史明[1] 王静[1] 黄建[1] 刘之荣[1] 

机构地区：[1]第四军医大学西京医院神经内科,西安710032

出　　处：《神经解剖学杂志》2014年第2期141-145,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81070950)

摘　　要：目的：对少突胶质前体细胞（oligodendrocyteprecursor cell，OPC）的传代方法进行改良，以建立一种低成本高效获取大量高纯度OPC的培养方法。方法：新生SD大鼠大脑皮层混合胶质细胞原代培养，待到细胞长满约65～75％培养瓶底，每日更换改良OPC生长培养基以刺激OPC的增殖，5～7d后OPC的密度足以进行传代时，采用EDTA化学分离为基础的机械振荡法获取oPc，并与传统的振荡分离法以及近年提出的单用化学分离法进行对比。结果：通过EDTA化学分离为基础的机械振荡法分离所得的OPC纯度明显高于另外两种分离方法，且数量显著多于传统的培养方法。结论：采用EDTA化学分离为基础的机械振荡改良传代法能高效获取大量高纯度体外培养的OPC。Objective： To set up an economical but efficient approach to obtain a large number of highly purified oligo- dendrocyte precursor cells （OPCs） via modification of the methods in OPC generation passage. Methods： The primary mixed glial cells from neonatal SD rat cerebral tissue were cultured, when the cultured cell mixture reached 65 -75% confluence, culture medium was replaced with modified OPC growth-medium （mOGM） to stimulate the proliferation of OPC and exchanged with fresh mOGM daily. At 5 -7 days, cultured OPCs reached a yield high enough for isolation and purification, then EDTA chemical-based separation followed with mechanical oscillation method were used for OPC obtai- ning. The quantity and purity of the OPC collected was compared with the traditional oscillation separation process and chemical separation only which was put forward in recent years. Results ：The purity of OPCs collected by EDTA chemical- based separation followed with meehanieal oscillation is significantly higher than the other two methods of passage, and the quantity is also significantly much more than the traditional culture methods. Conclusion： Using modified generation pas- sage approach of EDTA chemical-based separation followed with mechanical oscillation can efficiently obtain a large number of high purity of OPC cultured in vitro.

关 键 词：少突胶质前体细胞(OPC) 传代 EDTA 化学分离 振荡 纯度 大鼠 

分 类 号：R739.4[医药卫生—肿瘤]