从小鼠脂肪组织中分离纯化雪旺细胞的实验研究

目的 探索从新生小鼠脂肪组织中分离纯化雪旺细胞的可行性.方法 取新生(出生5～7 d)绿色荧光蛋白(GFP)转基因小鼠的腹股沟皮下脂肪,采用0.2%的复合胶原酶NB4消化法分离获取细胞,然后用含10%胎牛血清(FBS)的低糖DMEM培养基培养.用0.1%的复合胶原酶NB4差速分离纯化雪旺细胞,每48 h纯化1次,共进行2次纯化.用p75免疫荧光法鉴定雪旺细胞并分析其纯度.结果 从小鼠腹股沟皮下脂肪中分离纯化获得的细胞是雪旺细胞,经过2次纯化,其纯度可达90%.结论 从小鼠脂肪组织中可以分离纯化获得雪旺细胞,为组织工程人工神经种子细胞提供一种新的来源.

An experimental study on isolation and purification of Schwann cells from newborn mouse fat tissues

Objective To explore the possibility of isolating and purifying Schwann cells from newborn mouse fat tissues. Methods Adipose tissue was harvested from 5 to 7 day-old neonatal GFP mice. The tissue was digested with 0.2% compound collagenase NB4 to obtain cells. The derived cells were then cultured in lowglucose DMEM with 10% FBS for 24 hours. 0.1% compound collagenase NB4 was used to purify Schwann cells by differential isolation once every 48 hours for two times total. p75 fluorescent immunostaining was performed to determine the purity of Schwann cells. Results The cells isolated from mouse groin subcutaneous fat tissue were Schwann cells. The purity of Schwann cells reached as high as 90% after two time purification.Conclusion Purified Schwann cells can be isolated from mouse fat tissue, which provides a new source for seed cells for tissue-engineered nerves.