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目的:目前研究发现,周围神经中许旺细胞的标志物有很多,S100是其中之一。S100在体内表达的变化规律已有比较深入的研究,但是其在体外培养的许旺细胞的表达规律尚不清楚。因此,本课题研究小鼠许旺细胞在体外培养过程中S100蛋白的表达变化规律。方法:取新生(出生5-7 d)C57BL/6小鼠的坐骨神经,酶消化分离获取细胞后,培养纯化扩增3次。用S100免疫荧光法及RT-PCR技术研究许旺细胞在体外培养过程中S100的表达规律。结果:从坐骨神经消化所得到的许旺细胞,早期并不都表达S100,阳性率约为43.48%,随着培养时间延长(培养8天),所有许旺细胞均表达S100,能够达到阳性率95.66%。结论:体外培养的许旺细胞,其标志物S100阳性率表达随培养时间延长而增加。并且我们发现,S100并不能作为一个可靠的标志物来单独应用鉴定体外培养早期的许旺细胞。
Objective: The present study found that there are many markers of Schwann cells in the peripheral nerve, S100 is one of them. The variation of S100 expression in vivo has been studied in more depth, but its expression in Schwann cells cultured in vitro is unclear. Therefore, the subject of mouse Schwann cells in vitro culture S100 protein expression changes. Methods: The sciatic nerve of newborn (5-7 days old) C57BL / 6 mice was digested with enzyme and isolated. The cells were cultured and purified three times. S100 immunofluorescence and RT-PCR techniques were used to study the expression of S100 in Schwann cells cultured in vitro. Results: Schwann cells derived from the sciatic nerve digestion did not express S100 in early stage, and the positive rate was about 43.48%. With the prolongation of culture time (cultured for 8 days), all Schwann cells expressed S100 and reached a positive rate of 95.66 %. Conclusion: The expression of S100 in Schwann cells cultured in vitro increased with the prolongation of culture time. And we found that S100 can not be used as a reliable marker alone to identify early Schwann cells cultured in vitro.