新生大鼠少突胶质前体细胞的培养与鉴定

Isolation and identification of oligodendrocyte progenitor cells from neonatal rats

目的在体外培养条件下获取纯度较高的少突胶质前体细胞(oligodendrocyte progenitor cells,OPCs)。方法原代混合胶质培养,利用振荡法和差速贴壁法分离纯化少突胶质前体细胞,再用无血清化学条件培养基培养。免疫荧光法及膜片钳记录细胞电生理特性进行鉴定。结果振荡分离后超过90%细胞为NG2阳性OPCs;OPCs电生理特性:输入阻抗Ra相对较高;电压依赖的K+电流具有外向整流特性;对河豚毒素(tetrodotoxin,TTX)敏感的小的内向Na+流,不能产生动作电位。结论振荡法结合差速贴壁可以获得纯度较高的OPCs;OPCs具有区别于神经元、星型胶质细胞、小胶质细胞的独有电生理特性。

Objective To develop new methods to cultivate, retrieve and purify oligodendrocyte progenitor cells （OPCs）. Methods Primary OPCs were isolated from mixed glial cultures of newborn Sprague Dawley rat forebrains by using a selective detachment procedure, further purified by differential adhesion, and maintained in a chemical conditioned medium. The characteristic of OPCs was determined by both electrophysiological recording and immunostaining. Results Immunocytochemical analysis revealed that more than 90% of the isolated cells expressed the OPCs-specific marker NG2. OPCs exhibited a moderate input resistance, delayed rectifier K^＋ currents （Ik） that could be antagonized partly by TEA-Cl and CsCI, small tetrodotoxin （TTX） sensitive Na^ ＋ currents that failed to generate typical action potentials. Conclusion Isolation and purification by selective detachment and differential adhesion procedure are effective to obtain highly purified OPCs. OPCs, distinct from neuron, astrocyte, oligodendrocyte or microglia, possess unique physiological properties.