多发性骨髓瘤细胞条件培养液对人脐带间充质干细胞增殖和分化的影响  

Effects of multiple myeloma cell conditioned medium on proliferation and differentiation of human umbilical cord mesenchymal stem cells

作　　者：杨姁 王飞清 赵嘉宁 张弛可 梁惠玲 漆沙沙 吴丹 刘燕青 唐东昕 刘洋 李艳菊[3] Yang Xu;Wang Feiqing;Zhao Jianing;Zhang Chike;Liang Huiling;Qi Shasha;Wu Dan;Liu Yanqing;Tang Dongxin;Liu Yang;Li Yanju(Guizhou University of Traditional Chinese Medicine,Guiyang 550025,Guizhou Province,China;First Affiliated Hospital of Guiyang University of Traditional Chinese Medicine,Guiyang 550001,Guizhou Province,China;Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Key Laboratory of Adult Stem Cell Translational Research,Chinese Academy of Medical Sciences,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州中医药大学,贵州省贵阳市550025 [2]贵州中医药大学第一附属医院,贵州省贵阳市550001 [3]贵州医科大学附属医院,贵州省贵阳市550004 [4]中国医学科学院成体干细胞转化研究重点实验室,贵州省贵阳市550004

出　　处：《中国组织工程研究》2023年第10期1514-1520,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(31660326),项目负责人:刘洋;国家自然科学基金(82160519),项目负责人:李艳菊;国家自然科学基金后补助资金科研创新探索专项(2019YFC171250407),项目负责人:刘洋;国家自然科学基金后补助资金科研创新探索专项(2019YFC171250505),项目负责人:王飞清;中国博士后科学基金(2018M640938),项目负责人:刘洋;中国博士后科学基金(43XB3794XB),项目负责人:李艳菊;贵州省科学技术厅项目(黔科合基础[2016]1019),项目负责人:刘洋;贵州省科学技术厅项目(黔科合LH字[2017]7140号),项目负责人:刘燕青。

摘　　要：背景:近年来,在肿瘤微环境中干细胞与肿瘤的相互作用已成为研究热点,但是多发性骨髓瘤细胞条件培养液对人脐带间充质干细胞影响的实验鲜有报道。目的:探讨多发性骨髓瘤细胞条件培养液对人脐带间充质干细胞增殖、迁移及分化能力的影响。方法:以组织块贴壁法提取人脐带间充质干细胞,同时培养并提取人多发性骨髓瘤RPMI8226和U266细胞上清液制备条件培养液。按培养基的不同进行细胞分组:对照组人脐带间充质干细胞仅用L-DMEM完全培养液培养;实验组人脐带间充质干细胞用20%RPMI8226细胞条件培养液或20%U266细胞条件培养液和80%L-DMEM完全培养液培养。培养24 h,通过CCK-8、EDU、结晶紫染色观察细胞增殖情况,划痕法评估细胞迁移情况,成骨成脂诱导实验检测细胞分化能力,Real-time PCR检测细胞中周期蛋白基因p16、p21 mRNA表达。结果与结论:实验组细胞的增殖数量多于对照组(P <0.05),迁移率高于对照组(P <0.05),矿化结节染色面积小于对照组(P <0.05),脂滴阳性染色面积大于对照组(P <0.05);实验组的细胞周期基因p16、p21 mRNA表达低于对照组(P <0.05)。结果表明,人多发性骨髓瘤细胞条件培养液可促进人脐带间充质干细胞的增殖、迁移和成脂分化,抑制成骨分化,可能与其抑制p16、p21基因表达有关。BACKGROUND:In recent years,the interaction between stem cells and tumor in tumor microenvironment has become a hot topic.While few studies have reported the experiments of human umbilical cord mesenchymal stem cells in conditioned culture medium of human multiple myeloma cells.OBJECTIVE:To investigate the effects of conditioned medium of human multiple myeloma cells on proliferation,migration and differentiation of human umbilical cord mesenchymal stem cells.METHODS:Human umbilical cord mesenchymal stem cells were extracted by tissue block adherent method.At the same time,the supernatant of human multiple myeloma RPMI8226 and U266 cells was cultured and extracted to prepare conditioned culture medium.The cells were grouped according to different media:the control group was only cultured with L-DMEM complete culture medium.In experimental group,human umbilical cord mesenchymal stem cells were cultured with 20% RPMI8226 cell conditioned medium or 20% U266 cell conditioned medium and 80% L-DMEM complete medium.After 24 hours of culture,the proliferation of cells was analyzed by CCK8 assay,EDU,and crystal violet staining.The migration of cells was evaluated by the scrape method.Osteogenic and adipogenic induction experiments were used to detect cell differentiation.Real-time PCR was used to detect the mRNA expressions of cyclin genes p16 and p21 in cells.RESULTS AND CONCLUSION:The proliferation of cells in the experimental group was more than that in the control group(P < 0.05);the migration rate was higher than that in the control group(P < 0.05);the staining area of mineralized nodules was smaller than that of the control group(P < 0.05);and the positive staining area of lipid droplets was larger than that of the control group(P < 0.05);the mRNA expression of cell cycle genes p16 and p21 in the experimental group was lower than that in the control group(P < 0.05).These findings exhibit that conditioned culture medium of human multiple myeloma cells can promote the proliferation,migration,and adipogenic differentiat

关 键 词：多发性骨髓瘤 条件培养液 人脐带间充质干细胞 增殖 分化 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R394.2