体外自组装IKVAV纳米纤维凝胶在二维体系中对神经祖细胞分化的影响

Effect of self-assembling nanofiber gel containing IKVAV in two-dimensional culture system on differentiation of neural progenitor cells in vitro

背景：大量实验证明体外自组装的纳米支架材料可以促进神经祖细胞增殖与分化。目的：观察体外自组装含IKVAV纳米纤维凝胶在二维体系中对神经祖细胞分化的影响。方法：培养SD大鼠乳鼠神经祖细胞并用免疫荧光方法鉴定。在DMEM/F12触发下,含IKVAV的两亲性多肽溶液形成三维多孔凝胶,于透射电镜下观察其结构。将神经祖细胞分别接种到1%含IKVAV的两亲性多肽溶液及0.1g/L多聚赖氨酸包被的盖玻片上,培养1,3,7d后采用神经丝蛋白、胶质纤维酸性蛋白检测其分化情况。结果与结论：培养出巢蛋白阳性细胞,并且能分化为神经丝蛋白阳性的神经元与胶质纤维酸性蛋白阳性的胶质细胞,证实为神经祖细胞;含IKVAV的两亲性多肽溶液自组形成凝胶,并在透射电镜下显示为纳米纤维,直径为7.0－8.0nm,长度为100－1500nm;IKVAV的两亲性多肽溶液组向神经元分化得能力明显优于多聚赖氨酸组。说明体外自组装IKVAV纳米纤维凝胶在二维培养体系中对神经祖细胞分化有一定的促进作用。

BACKGROUND：Numerous studies have demonstrated that the self-assembling nanofiber promotes neural progenitor cells proliferation and differentiation.OBJECTIVE：To study the differentiation of neural progenitor cells on self-assembling nanofiber gel containing IKVAV in two-dimensional culture system in vitro.METHODS：Neural progenitor cells of SD neonatal rats were cultured in culture medium and examined by immunofluorescence.Amphipathic polypeptide solution containing 1% IKVAV-PA was trigger to form three-dimensional porous gel by the addition of DMEM/F12,and the structure was detected under transmission electron microscope.Neural progenitor cells were seeded on the surface of coverslips covered with 1% KVAV-PA or covered with 0.1 g/L polylysine,and the cell differentiation was determined by neurofilament and glial fibrillary acidic protein after cultivated 1,3,7 days respectively.RESULTS AND CONCLUSION：The cultured Nestin-positive cells were found,and they were able to differentiated into neurofilament-positive neurons and glial fibrillary acidic protein-positive glial cells,confirming as neural progenitor cells;Transmission electron microscopy showed that the gel derived from amphipathic polypeptide solution containing IKVAV-PA,was composed of nanofiber network,their diameter ranging 7.0-8.0 nm and the length 100-1 500 nm.The ability of differentiation into neurons of the amphipathic polypeptide solution containing IKVAV-PA surpasses to the polylysine.The self-assembling nanofiber gel contained IKVAV in two-dimensional culture system may promote the differentiation of neural progenitor cells in vitro.