脂肪干细胞上清液培养雪旺细胞的实验研究

An Experimental Study on Cultivating Schwann Cells in the Supernatant of the Culture Medium of Adipose-derived Stem cells(ADSC-CM)

目的:研究应用脂肪干细胞上清液培养新生小鼠雪旺细胞的可行性。方法:取新生(出生5-7天)C57BL/6小鼠的坐骨神经,采用0.2%的复合胶原酶NB4消化法分离获取细胞,然后应用雪旺细胞条件培养基(SCCM)和C57BL/6小鼠的脂肪干细胞上清液(ADSC-CM)分别培养雪旺细胞。用0.2%复合胶原酶NB4差速分离纯化这两种方法培养的雪旺细胞,每48 h纯化1次,共进行2次纯化。应用P75免疫荧光染色方法鉴别两组P2代雪旺细胞并比较两组雪旺细胞的纯度和生长情况。结果:脂肪干细胞上清液培养的雪旺细胞纯化两次后,数量明显增多,其纯度与雪旺细胞条件培养基相比没有明显差异(P>0.05)。结论:脂肪干细胞上清液可以较好的培养雪旺细胞,可以作为一种新的廉价方便的培养基代替雪旺细胞条件培养基来培养许雪旺细胞。

Objective： To explore the possibility of cultivating newborn mouse＇s schwann cells in the supematant of the culture medium of Adipose-derived stem cells （ADSC-CM）. Methods： Sciatic nerves were harvested from 5 to 7 day-old C57BL/6 mice.Sciatic nerves were digested with 0.2%compound collagenase NB4 to obtain schwann cells. Schwann cells were cultured in two culture medium, one is conditional medium of Schwann cells （SCCM）, the other is supematant of the culture medium of Adipose-derived stem cells （ADSC-CM）. 0.2% compound collagenase NB4 was used to purify these schwann cells by differential isolation once every 48 hours for two times total. P75 fluorescent immunostaining was performed to determine the purity and growth situation of schwann cells. Results： After two times purification, the numbers of these schwarm cells in ADSC-CM were significantly improved, and the differences of purity of these two methods cultured schwann cells were not statistically significant （P〉0.05）. Conclusion： ADSC-CM can have a good effect on culture schwann cells. As a new cheap convenient culture medium, it can replace SCCM to culture schwann cells.