Jin Yu-Qing, Liu Wei, Hong Tan-Hui, Cao Yilin
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Tissue Engineering Laboratory, Shanghai, China.
J Neurosci Methods. 2008 May 15;170(1):140-8. doi: 10.1016/j.jneumeth.2008.01.003. Epub 2008 Jan 16.
Schwann cell purification is usually difficult due to the contamination of fibroblasts, which often become a predominant cell type in Schwann cell culture in vitro. We have developed a novel and efficient method to enrich Schwann cells by differential detachment of two types of cells. In the culture, cells were treated with a multiplex collagenase and the Schwann cells were found to detach faster than fibroblasts and thus Schwann cells could be easily isolated. Within 5 days, Schwann cell purity could reach above 99%, which was confirmed by immunostaining characterization and flow cytometric analysis. In addition, this efficient method can reach a high cell yield after two rounds of differential detachment procedures and does not require antimitotic treatment or special equipment as often needed by other reported methods.
雪旺细胞的纯化通常很困难，因为会受到成纤维细胞的污染，而成纤维细胞在体外雪旺细胞培养中常常成为主要的细胞类型。我们开发了一种新颖且高效的方法，通过两种细胞的差异脱离来富集雪旺细胞。在培养过程中，细胞用复合胶原酶处理，发现雪旺细胞比成纤维细胞脱离得更快，因此雪旺细胞能够轻易地被分离出来。在5天内，雪旺细胞纯度可达到99%以上，这通过免疫染色鉴定和流式细胞术分析得以证实。此外，这种高效方法经过两轮差异脱离程序后可获得高细胞产量，并且不需要像其他报道方法通常所需要的抗有丝分裂处理或特殊设备。
