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为了探讨周围神经组织环境因素对EPI-NCSCs(epidermal neural crest stem cells)分化的影响,该文用坐骨神经组织提取液为环境因素,对体外培养的EPI-NCSCs进行分化诱导。以正常培养的EPI-NCSCs为对照组,细胞培养液成分为:DMEM/F12、10%FBS、2%B27、20 ng/mL bFGF。在细胞培养液中添加1%周围神经组织液作为实验组。MTT结果显示,周围神经组织液对EPI-NCSCs的活性和增殖无影响;对照组和实验组EPI-NCSCs都呈多种形态。RT-PCR分析显示,在对照组和实验组中EPI-NCSCs均表达GFAP、S-100、β-III Tubulin(Tuj1),但在实验组中,这3种蛋白的表达量均高于对照组。免疫组化分析显示,在对照组中,EPI-NCSCs同时表现了雪旺细胞和早期神经元的鉴定特征;在实验组中,EPI-NCSCs没有神经元的鉴定特征,只表达雪旺氏细胞的鉴定特征。这表明,周围神经组织液可以诱导体外培养的EPI-NCSCs向雪旺氏细胞分化。
In order to investigate the effect of environmental factors on the differentiation of epidermal neural crest stem cells (EPI-NCSCs), the sciatic nerve tissue extract was used as an environmental factor to induce the differentiation of EPI-NCSCs in vitro. The normal culture EPI-NCSCs as control group, cell culture fluid composition: DMEM / F12, 10% FBS, 2% B27, 20 ng / mL bFGF. 1% peripheral nerve tissue fluid was added to the cell culture medium as experimental group. MTT results showed that peripheral nerve tissue fluid had no effect on the activity and proliferation of EPI-NCSCs; EPI-NCSCs in the control group and experimental group showed various forms. RT-PCR analysis showed that GFAP, S-100 and β-III Tubulin (Tuj1) were all expressed in EPI-NCSCs in the control and experimental groups, but in the experimental group, the expression of these three proteins was higher than that in the control group . Immunohistochemical analysis showed that in the control group, EPI-NCSCs simultaneously demonstrated the identification characteristics of Schwann cells and early neurons. In the experimental group, EPI-NCSCs did not identify the neurons and only expressed Schwann cells Characterization. This indicates that peripheral nerve tissue fluid can induce EPI-NCSCs cultured in vitro to differentiate into Schwann cells.