Flores-Obando Rafael E, Freidin Mona M, Abrams Charles K
Program in Molecular and Cellular Biology, State University of New York Downstate Medical Center.
Department of Neurology and Rehabilitation, University of Illinois at Chicago.
J Vis Exp. 2018 May 21(135):57543. doi: 10.3791/57543.
The efficient and robust isolation and culture of primary oligodendrocytes (OLs) is a valuable tool for the in vitro study of the development of oligodendroglia as well as the biology of demyelinating diseases such as multiple sclerosis and Pelizaeus-Merzbacher-like disease (PMLD). Here, we present a simple and efficient selection method for the immunomagnetic isolation of stage three O4 preoligodendrocytes cells from neonatal mice pups. Since immature OL constitute more than 80% of the rodent-brain white matter at postnatal day 7 (P7) this isolation method not only ensures high cellular yield, but also the specific isolation of OLs already committed to the oligodendroglial lineage, decreasing the possibility of isolating contaminating cells such as astrocytes and other cells from the mouse brain. This method is a modification of the techniques reported previously, and provides oligodendrocyte preparation purity above 80% in about 4 h.
高效且稳健地分离和培养原代少突胶质细胞（OLs）是体外研究少突胶质细胞发育以及脱髓鞘疾病（如多发性硬化症和佩利措伊斯-梅茨巴赫样病（PMLD））生物学特性的宝贵工具。在此，我们展示一种简单有效的筛选方法，用于从新生小鼠幼崽中免疫磁珠分离三期O4少突胶质前体细胞。由于在出生后第7天（P7），未成熟的OL占啮齿动物脑白质的80%以上，这种分离方法不仅能确保高细胞产量，还能特异性分离已确定为少突胶质细胞谱系的OL，降低从小鼠脑中分离污染细胞（如星形胶质细胞和其他细胞）的可能性。该方法是对先前报道技术的改进，约4小时内可提供纯度高于80%的少突胶质细胞制剂。
