中国组织工程研究Issue(42)：7420-7426,7.DOI:10.3969/j.issn.2095-4344.2013.42.013
季铵盐壳聚糖三维支架复合GNDF载间充质干细胞向神经样细胞分化
Neuron-like differentiation of mesenchymal stem cells induced by quaternary chitosan thermosensitive hydrogel scaffolds combined with glial cell line-derived neurotrophic factor
摘要
Abstract
BACKGROUND:In recent years, chitosan-based thermosensitive hydrogel, as scaffold materials, have received more and more attentions in the field of tissue repair because of good biocompatibility, biodegradability and drug-sustained release. <br> OBJECTIVE:To explore the directed differentiation and growth of rat bone marrow mesenchymal stem cells on the quaternary chitosan thermosensitive hydrogel scaffold and to look for more ideal tissue engineering materials for the treatment of nervous system damage. <br> METHODS:The thermosensitive hygrogel scaffold was prepared using hydroxypropyltrimethyl ammonium chloride chitosan (HACC) andβ-glycerophosphate (β-GP). The spatial structure of scaffold was observed by scanning electronic microscope. Effect of leaching liquor from the HACC/β-GP scaffold on the viability of bone marrow mesenchymal stem cells was detected by (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The albumin from bovine serum was combined with the scaffold, and the slow-release effect of the scaffold was detected by ultraviolet absorption spectrometry. Bone marrow mesenchymal stem cells were incubated onto the compound scaffold at 3 passages. The adhesion, growth and differentiation of bone marrow mesenchymal stem cells on the compound scaffold were observed by the scanning electron microscope. Neuron-specific enolase was detected by immunofluorescence. <br> RESULTS AND CONCLUSION:The porosity and thermal sensitivity of HACC/β-GP scaffold and slow-release effect of glial cellline-derived neurotrophic factor were apparent. The results of MTT showed that the compound scaffold cannot take apparent negative effects to the proliferation of bone marrow mesenchymal stem cells. After inoculation, bone marrow mesenchymal stem cells permeated the porous structure of the scaffold and adhered to the scaffold. Under the role of glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells showed neuron-like cellmorphology and cells co-cultured with the compound scaffold expressed the marker of neurons, neuron-specific enolase. Under the role of slow-release glial cellline-derived neurotrophic factor, bone marrow mesenchymal stem cells can grow wel in vitro and differentiate into neuron-like cells on the HACC/β-GP scaffold.关键词
生物材料/生物材料基础实验/组织工程神经材料/季铵盐壳聚糖/细胞支架/骨髓间充质干细胞/细胞培养/胶质细胞源性神经营养因子/分化/神经元/神经元烯醇化酶/国家自然科学基金Key words
chitosan/biocompatible materials/mesenchymal stem cells/neurons/tissue engineering分类
医药卫生引用本文复制引用
黄成,杨建东,冯新民,李广峰,李艺楠,肖海祥,孙钰..季铵盐壳聚糖三维支架复合GNDF载间充质干细胞向神经样细胞分化[J].中国组织工程研究,2013,(42):7420-7426,7.基金项目
国家自然科学基金面上项目《载胶质细胞源性神经营养因子基因的壳聚糖复合骨髓间充质干细胞修复脊髓损伤》提供科研经费资助，项目编号81071466@@@@the National Natural Science Foundation of China, No.81071466 （）
