改良的酶消化后植块法体外培养雪旺细胞

Culturing Schwann cells by explants and modified enzyme digestion technique

目的 观察改良的酶（浓度为2g/L的胶原酶）消化后植块法体外培养雪旺细胞（SCs）效果,探讨高效获取高纯度、高活性SCs的培养纯化方法.方法 取新生SD大鼠坐骨神经,剥离神经外膜,剪块,分别采用单酶消化后植块法与双酶消化法进行培养.以活细胞计数和S-100单抗标记相结合判断SCs增殖和纯化程度,比较2种方法的优劣.结果 改良的单酶消化后植块法可获取3.5x106个SCs,成活率为96%,纯度达94%以上：双酶消化法约获取3.0×106个SCs,成活率为92%,纯度达90%.结论 采用改良的酶消化后植块法培养SCs,可较快得到数量多、纯度高的SCs,符合实验要求,值得推广应用.

Objective To introduce an efficient method for culturing and purifying Schwann cells （SCs） in vitro. Methods Sciatic nerves were harvested from neonatal SD rats and the epineuria were removed. Single enzyme digestion combined with explans and double enzyme digestion were employed, respectively, to digest the nerve tissues following trituration. The proliferation of SCs and degree of purification were evaluated by viable count method and the combination of S-100 labeled monoclonal antibody and SCs. Results Proximally 3.5 × 106 cells were harvested with 96% survival rate and a purity of Schwann cells over 94% by single enzyme digestion combined with explans, however only 3.0×106 cells were gained with the purity being 90% and survival rate being 92% by double enzyme digestion. Conclusion This method yields large amount of viable Schwann cells with high purity and survival rate.