School of Biology and Basic Medical Science, Suzhou University, Suzhou, Jiangsu 215006, P.R. China.
Jiangsu Key Laboratory of Neuroregeneration, Co‑innovation Center of Neuroregeneration, Nantong University, Nantong, Jiangsu 226001, P.R. China.
Mol Med Rep. 2017 Aug;16(2):1747-1752. doi: 10.3892/mmr.2017.6777. Epub 2017 Jun 14.
The present study presented a protocol that can be used to obtain rapidly a high purity of proliferating rat Schwann cells from freshly dissociated rat peripheral nerves. The sciatic nerves of newborn rats (1‑3 day old) were dissociated, and the Schwann cells (SCs) were purified using fluorescence‑activated cell sorting (FACS) based on the SC membrane‑specific expression of the low‑affinity nerve growth factor receptor, p75NGFR and oligodendrocyte marker 4. Following sorting, the cells were plated on poly‑l‑lysine‑coated dishes in SC culture medium containing DMEM with 10% FBS, 1% penicillin/streptomycin, 2 µM forskolin and 10 ng/ml HRG. The purified rat SCs were propagated for passaging until confluent. This protocol resulted in SC cultures, which were >98% pure. This FACS‑based protocol can be used to facilitate future investigations of general SC biology.
本研究提出了一种方案，可用于从新鲜分离的大鼠周围神经中快速获得高纯度增殖的大鼠雪旺细胞。取新生大鼠（1-3 日龄）的坐骨神经进行解离，然后根据低亲和力神经生长因子受体 p75NGFR 和少突胶质细胞标志物 4 在雪旺细胞膜上的特异性表达，使用荧光激活细胞分选（FACS）对雪旺细胞（SCs）进行纯化。分选后，将细胞铺在涂有多聚赖氨酸的培养皿中，置于含有 10% FBS、1%青霉素/链霉素、2µM forskolin 和 10ng/ml HRG 的 DMEM SC 培养基中。纯化的大鼠雪旺细胞传代培养至汇合。该方案得到的雪旺细胞培养物纯度>98%。这种基于 FACS 的方案可用于促进未来对雪旺细胞一般生物学的研究。
