猪胎儿神经干细胞的分离培养和分化

摘要

本研究旨在从猪胎儿脑组织中分离培养神经干细胞,观察神经干细胞生长特性和体外增殖、分化特点.利用神经干细胞培养体系,从胎龄30 d的猪胎儿脑组织中分离培养神经干细胞并诱导神经干细胞贴壁分化,采用RT-PCR技术检测干细胞和分化细胞表面标志或相关基因.结果成功分离培养出神经干细胞,神经干细胞具有分化潜能.神经干细胞中Nestin表达强阳性,β-actin、DCX、Hesl、Oct4、Desmin、CD-90、Nanog和Sox2表达阳性;体外诱导的神经干细胞可以分化为星形胶质细胞(表达GFAP)、少突胶质细胞(表达GalC)和神经元细胞(表达NF、NSE和MAP2).结果提示,从猪胎儿脑组织分离神经干细胞具有可行性和有效性,神经干细胞具有自我更新、增殖和分化潜能.%This experiment was conducted to study the charicteristics of porcine neural stem cells. Brains were dissected from 30-day porcine fetuses, enzymatically dissociated, and grown in the presence of epidermal growth factor, basic fibroblastgrowth factor etc. Neural stem cells were in-duced to differentiate in vitro. Expanded neural stem cells and differentiated cells were harvested for analysis using reverse transcription-polymerasechain reaction (RT-PCR). Porcine neural stem cells could be grown as suspended spheres, and they expressed Nestin, β-actin, DCX, Hesl, Oct4, Desmin, CD-90, Nanog and Sox2. They were differentiated into astrocyte (GFAP), oligo-dendrocyte (GalC) and neuron (NF,NSE and MAP2). This study shows that porcine neural stem cells can be cultured over the long term and are able to self-renew and differentiate into neurons, astrocytes, and oligodendroeytes in vitro.