神经干细胞体外三维培养模型的构建  

Establishment of neural stem cells in vitro three-dimensional model

作　　者：关水[1] 葛丹[1] 陆瑞欣[1] 刘天庆[1] 马学虎[1] 崔占峰 

机构地区：[1]大连理工大学干细胞与组织工程研究室,辽宁大连116024 [2]Oxford Centre for Tissue Engineering and Bioprocessing,Department of Engineering Science,Oxford University,Oxford OX13PJ,U.K.

出　　处：《中国科技论文在线》2011年第3期181-186,共6页

基　　金：国家自然科学基金资助项目(30800288);高等学校博士学科点专项科研基金资助项目(20070141050);大连理工大学科研启动基金资助项目(1000893364)

摘　　要：研究了小鼠海马NSCs在胶原凝胶内的生长情况,构建适合于干细胞药物筛选及作用机理研究的三维培养模型。实验采用体外培养第三代NSCs,均匀混合于胶原溶液中,形成"细胞-胶原"的三维凝胶培养。通过扫描电镜观察胶原的微观结构,倒置显微镜及激光共聚焦显微镜观察胶原凝胶中NSCs的生长、伸展情况;应用免疫荧光技术鉴定NSCs,同时采用CCK-8法检测不同胶原浓度及不同培养模式下NSCs的生存和增殖能力。结果表明:质量浓度为0.5 mg/mL的胶原具有良好的三维多孔结构,凝胶中的NSCs生长状态良好,且大部分呈Nestin、BrdU阳性;与传统二维培养方式相比,三维胶原凝胶更利于NSCs的生存及增殖。Neural stem cells（NSCs）,obtained from the mouse hippocampus,were cultured in the three-dimensional（3D） collagen gel in order to build a suitable 3D culture model for the studies of NSCs drug screening and mechanism.The ＂cell-collagen＂ 3D gel was formed by mixing NSCs of the third generation into the collagen solution.The microstructure of collagen was observed by scanning electron microscopy and the growth and stretching of NSCs in collagen gel were detected by inverted microscope and laser scanning confocal microscope.And,we applied the immunofluorescence techniques to identify NSCs and used the CCK-8 method to detect the survival and proliferation of NSCs in different concentrations of collagen and under different culture modes.The results showed that the mass concentration of 0.5 mg/mL collagen has a right 3D porous structure;NSCs in gel grow well and most are Nestin,BrdU-positive;compared with the traditional two-dimensional（2D） modes,3D collagen gel is more conducive to the survival and proliferation of NSCs.

关 键 词：神经干细胞 三维培养 胶原凝胶 

分 类 号：Q421[生物学—神经生物学]