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目的:体外原代培养脂肪源性干细胞(adipose-derived stem cell,ADSC),并在细胞水平检测各种神经营养因子(neurotrophic factor,NF)的表达情况,探讨ADSC是否可以作为神经损伤修复的种子细胞。方法:体外原代培养和鉴定ADSC,应用RT-PCR方法检测ADSC细胞神经营养因子,包括神经生长因子(nerve growth factor,NGF)、脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、神经营养素-3(neurotrophin 3,NT-3);免疫组织化学法检测粘附因子(nerve cell adhesion molecule,NCAM)表达情况。结果:ADSC可以向脂肪和骨两方面分化;RT-PCR结果显示ADSC细胞NGF、BDNF、NT-3 mRNA水平明显升高;同时免疫荧光法检测到了高水平的NCAM。结论:ADSC具有干细胞分化潜能,并且能够表达NF和NCAM。
OBJECTIVE: To primarily culture adipose-derived stem cells (ADSCs) in vitro and to detect the expression of various neurotrophic factors (NFs) at the cellular level to investigate whether ADSCs can be used as seeds for repair of nerve injury cell. Methods: ADSCs were cultured and identified in primary culture in vitro. RT-PCR was used to detect neurotrophic factors, including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) Neurotrophin 3 (NT-3) was detected by immunohistochemistry. The expression of nerve cell adhesion molecule (NCAM) was detected by immunohistochemistry. Results: ADSCs could differentiate into adipose and bone. RT-PCR results showed that the expression of NGF, BDNF and NT-3 mRNA in ADSCs was significantly increased. Meanwhile, high level of NCAM was detected by immunofluorescence. Conclusion: ADSC has stem cell differentiation potential and can express NF and NCAM.