改良冷喷注法纯化大鼠雪旺细胞的实验研究

Purification of Schwann cells with improved Cold Jet method

目的报道一种新的雪旺细胞提纯方法一改良冷喷注法，并与其他2种方法进行对比，评价其优缺点。方法分离新生6d的Wistar大鼠双侧坐骨神经，双酶消化法分离雪旺细胞，用S-100免疫细胞化学染色方法对雪旺细胞进行鉴定。将收集到的细胞分为3组，分别应用冷喷注法、改良冷喷注法和酶消化法对细胞进行纯化，计算纯化后的细胞纯度，流式细胞仪测定细胞活性。结果改良冷喷注法提纯出的雪旺细胞，纯度和细胞活性与冷喷注法比较差异无统计学意义（P〉0．05），但二者均高于酶消化组，差异有统计学意义（P〈0．05）。结论改良冷喷注法提纯雪旺细胞的优点是操作简便，获得雪旺细胞纯度高且不影响细胞活性，缺点是对操作者实验操作技术和经验要求相对较高，初学者不易掌握。

Objective To evaluate the advantages and disadvantages of improved Cold Jet method, a new method to separate Schwann cells （SCs） through comparing it with the other 2 methods （double enzyme digestion and Cold Jet method）. Methods SCs were obtained fTom sciatic nerves of 6-d-old Wistar neonatal rats with the method of double enzyme digestion; S-100 immunocytochemistry was used to identify the SCs. These cells were divided into 3 groups and 3 different methods were used to purify the cells, separately. After that, we calculated the purity by morphology method, and flow cytometry was employed to examine the cell viability. Results No statistical significance on the purification and cell viability was noted between cells obtained with improved Cold Jet method and cells obtained with Cold Jet method （P〉0.05）, but cells fi＇om both methods enjoyed significantly higher levels of purification and cell viability than cells purified with enzyme digestion （P〈0.05）. Conclusion The improved Cold Jet method is simple in operation and can obtain high purification of SCs with good cytoactive; however, it needs skillful technique and lots of experiments.