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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 study the feasibility of culturing newborn mouse Schwann cells with adipose-derived stem cell supernatant. METHODS: The sciatic nerve of newborn (5-7 days old) C57BL / 6 mice was isolated and harvested by using 0.2% collagenase NB4 digestion method. Then Schwann cell conditioned medium (SCCM) and C57BL / 6 mice Schwann cells were cultured in adipose-derived stem cell supernatant (ADSC-CM). The Schwann cells cultured in these two methods were separated and purified by 0.2% complex collagenase (NB4), and purified once every 48 hours. P75 immunofluorescence staining method was used to identify two groups of P2 generation of Schwann cells and compare the purity and growth of two groups of Schwann cells. Results: Schwann cells cultured in adipose-derived stem cell supernatant were purified twice and the number of Schwann cells increased obviously. The purity of Schwann cells was not significantly different from that of Schwann cells (P> 0.05). CONCLUSION: Adipose-derived stem cell supernatant can culture Schwann cells better, and can be used as a new inexpensive and convenient medium to replace Schwann cell conditioned medium to culture Schwann cells.