利用藻酸盐与骨髓间充质干细胞及前列腺癌细胞混合培养:观察干细胞对癌细胞增殖速度及成簇大小的影响  

Co-culture of prostate carcinoma cells with alginate and bone marrow mesenchymal stem cells:To observe the effect of stem cells on proliferation speed and clustering size of prostate carcinoma cells

作　　者：解杰[1] 陈安民[1] 郭风劲[1] 王建超[1] 廖晖[1] 柳昊[1] 陈超[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,湖北省武汉市430030

出　　处：《中国组织工程研究与临床康复》2010年第6期1009-1014,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：课题受国家973重点基础研究发展规划资助项目(2002CB513107);教育部新教师基金项目(200804871051)资助~~

摘　　要：背景:肿瘤外环境研究的模型主要集中在实验室细胞的二维培养与体外动物实验,缺乏三维立体模型的研究与构建。目的:体外模拟肿瘤骨转移的三维模型,并比较在有无骨髓间质干细胞时前列腺癌细胞的增殖速度与成簇大小以及成簇率。方法:清洁级SD大鼠2只用于提取骨髓间质干细胞。利用藻酸盐模拟骨髓微环境,将前列腺癌细胞与SD大鼠骨髓间质干细胞在藻酸钙中共培养。通过显微镜与组织切片观察细胞在模型中的生长情况。利用绿色荧光蛋白转染肿瘤细胞,通过培养在普通光及荧光显微镜下观察三维模型中肿瘤单克隆的生长扩增情况。结果与结论:有干细胞存在共培养的三维藻酸钙微环境中前列腺癌细胞的成簇速度、数量及成瘤率均较对照组高。对照组在7.75d、有干细胞的混合培养组在6.00d能形成单克隆细胞簇,10d后对照组瘤簇细胞计数为(95.13±11.63)个;有干细胞的混合培养组为(112.53±14.67)个,荧光细胞克隆形成率对照组为(77.10±6.85)%;有干细胞的混合培养组为(64.55±6.21)%;两者相比均有显著性意义。结果提示前列腺癌细胞与骨髓基质干细胞在藻酸钙微球环境中,有利于细胞增殖及成簇性发展。BACKGROUND：Models concerning tumor external environment mainly concentrated on laboratory two-dimensional culture and in vitro animal experiment,which lack of three-dimensional stereo.OBJECTIVE：To establish in vitro bone metastasis stereo models of human prostate carcinoma,and to investigate the effect of stem cells on proliferation rate and clustering size of prostate carcinoma cells.METHODS：Bone marrow mesenchymal stem cells （BMMSCs） were extracted from 2 clean grade SD rats.Alginate was used to simulate medullary microenvironment,where prostate carcinoma cells and BMMSCs were co-culturedd.Growth of the cells in the three-dimensional model was observed through microscope and histological sections.The carcinoma cells were transfected with green fluorescent protein.The proliferation of monoclonal cells clustering was observed under light microscope and fluorescence microscope.RESULTS AND CONCLUSION：In the co-culture group,the clustering speed,clustering amount and tumor formation rate were greater that those of the control group.The monoclonal cells clustering was formed at 7.75 days and 6.00 days in the control and co-culture groups,respectively,with cell counts of （95.13±11.63） and （112.53±14.67） after 10 days.The formation rate of fluorescent cell clones was （77.10±6.85）% in the control group and （64.55±6.21）% in the co-culture group,the difference had significance.The results suggested that：the alginate microenvironment is conductive to proliferation and clustering of prostate carcinoma cells and BMMSCs.

关 键 词：前列腺癌 骨转移模型 共培养 藻酸钙凝胶 绿色荧光标记 骨髓间充质干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]