多肽二维凝胶支架材料与鼠神经干细胞的细胞相容性

Cytocompatibility of Mouse Neural Stem Cells Cultivated in the Surface of the Two-dimensional Scaffold of the Polypeptide Hydrogel

目的研究鼠神经干细胞在二维自组装多肽凝胶支架材料表面生长及分化情况,证明多肽凝胶可作为神经组织工程支架材料。方法取新生1周内乳鼠的大脑皮质,机械分离出神经干细胞,无血清培养液(DMEM/F12+bFGF+EGF+B27)培养,免疫组化SABC法对神经干细胞进行鉴定;然后,将其接种到凝胶支架材料表面(实验组)及多聚赖氨酸包被的盖玻片表面(对照组),倒置相差显微镜观察干细胞的生长及分化情况。结果免疫组化法鉴定:培养的细胞为神经干细胞,Nestin(+)。神经干细胞可分化为神经元及星形胶质细胞,NSE(+),GFAP(+)。倒置相差显微镜观察:实验组中,接种的神经干细胞生长良好,7 d后可分化为有突起神经细胞;对照组中,7 d后未发现长出突起神经细胞。结论二维自组装多肽凝胶对神经干细胞有良好细胞相容性,可作为神经组织工程支架材料。

Objective Mouse neural stem cells （NSCs） growing and differentiatitig in the surface of the two-dimensional and self-assembly polypeptide hydrogel was investigated in order to testify that polypeptide hydrogel was characterized as the top scaffold for neural tissue engineering. Methods NSCs harvested from the cerebral cortex of neonatal one-week mice were triturated and cultivated in serum-free DMEM/F12 ＋ bFGF＋ EGF＋ B27, then identified by the immunohistochemistry SABC method. Subsequently NSCs were implanted in the surface of both the hydrogel （experimental group） and coverslip covered with polylysine （control group）, and the morphology of growth and differentiation of cells was demonstrated by inverted phase contrast microscopy. Results NSCs, which were differentiated into neuron and astrocyte stained with NSE（＋ ） and GFAP（＋ ） respectively, were stained with Nestin（＋ ）. Implanted NSCs in experimental group grew and extended process which was not found in control group after 7 days. Conclusion The two-dimensional polypeptide hydrogel was good cytocompatible to NSCs and was used as scaffold of neural tissue engineering.