Dugas Jason C, Emery Ben
Myelin Repair Foundation, Saratoga, California 95070, USA.
Cold Spring Harb Protoc. 2013 Aug 1;2013(8):745-58. doi: 10.1101/pdb.prot070862.
This protocol describes how to purify oligodendrocyte precursor cells (OPCs) from postnatal rodent brains. The method utilizes an immunopanning technique to first remove unwanted cells by negative selection and then purify OPCs by positive selection and subsequent enzymatic release from the final panning plate. Included are modifications that allow for purification and culturing of OPCs from mouse instead of rat tissue and for use of optic nerves instead of whole brains. The method for isolating OPCs from whole brain can be used for isolating OPCs from any specific region of the brain, provided that the area can be dissected away from the rest of the tissue. Suggested culture media for maintaining proliferating OPCs or inducing oligodendrocyte (OL) differentiation are also described.
本方案描述了如何从出生后的啮齿动物大脑中纯化少突胶质前体细胞（OPC）。该方法利用免疫淘选技术，首先通过阴性选择去除不需要的细胞，然后通过阳性选择并随后从最终的淘选板上酶解释放来纯化OPC。其中包括一些改进方法，可用于从小鼠而非大鼠组织中纯化和培养OPC，以及使用视神经而非全脑。从全脑中分离OPC的方法可用于从大脑的任何特定区域分离OPC，前提是该区域能够从其余组织中分离出来。文中还描述了用于维持增殖性OPC或诱导少突胶质细胞（OL）分化的推荐培养基。
