人骨髓CD106^+间充质干细胞亚群与CD106^-间充质干细胞亚群生物学功能的比较  

Comparison of the Biological Functions between Human Bone Marrow Derived CD106^+ Mesenchymal Stem Cells and CD106^- Subgroup

作　　者：卢士红 葛美丽 尤亚红 霍佳莉 梁昊岳 于文颖 杨栋林 冯四洲 韩忠朝 LU Shihong;GE Meili;YOU Yahong;HUO Jiali;LIANG Haoyue;YU Wenying;YANG Donglin;FENG Sizhou;HAN Zhongchao(State Key Laboratory of Experimental Hematology,Institute of Hematology and Blood Diseases Hospital,CAMS and PUMC,Tianjin 300020,China)

机构地区：[1]中国医学科学院北京协和医学院血液病医院血液学研究所国家重点实验室

出　　处：《中国医学科学院学报》2019年第4期443-451,共9页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(81330015、81730006、81300388)~~

摘　　要：目的探讨人骨髓间充质干细胞(MSCs)中CD106+MSCs亚群与CD106-MSCs亚群生物学功能的差异。方法取人骨髓扩增MSCs,然后分选CD106+MSCs亚群与CD106-MSCs亚群,检测细胞增殖、黏附功能、趋化功能、向成脂成骨分化能力、衰老情况、衰老蛋白p21;检测细胞受到肿瘤坏死因子-α(TNF-α)刺激时,胞内核因子-κB(NF-κB)转入细胞核内的能力等。结果 CD106+MSCs亚群比CD106-MSCs亚群增殖功能增强,CD106+MSCs亚群组光密度(OD)值明显高于CD106-MSCs亚群组OD值(48 h:1.004±0.028比0.659±0.023,t=3.946,P=0.0225;72 h:2.574±0.089比1.590±0.074,t=11.240,P=0.0000)。CD106+MSCs亚群组比CD106-MSCs亚群组黏附能力更强,两组OD值分别为0.648±0.018、0.418±0.0234,两组比较差异有统计学意义(t=7.869,P=0.0002)。CD106+MSCs亚群组趋化能力明显高于CD106-MSCs亚群组,其趋化细胞数分别为114.500±4.481、71.000±4.435,两组比较差异有统计学意义(t=6.900,P=0.0005)。CD106+MSCs亚群组较CD106-MSCs亚群组被诱导向成骨分化增强而向成脂分化减弱。CD106+MSCs亚群组衰老细胞和衰老蛋白p21的表达较CD106-MSCs亚群组更少,β-半乳糖苷酶染色检测P5代MSCs分选出的CD106+MSCs亚群组未见着色细胞,而CD106-MSCs亚群组则出现少量绿色着色细胞;其衰老蛋白p21在CD106+MSCs亚群组表达率明显低于CD106-MSCs亚群组[(17.560±1.421)%比(45.800±2.569)%,t=9.618,P=0.0000]。而用TNF-α刺激MSCs 0.5 h,CD106^+MSCs亚群组胞内NF-κB入核率明显高于CD106-MSCs亚群组[(37.780±3.268)%比(7.30±1.25)%,t=8.713,P=0.0001]。结论人骨髓CD106+MSCs亚群比CD106-MSCs亚群在骨髓微环境中展示了更强的生物学功能活力。Objective To analyze the differences in biological functions between bone marrow(BM)-derived CD106+mesenchymal stem cells(MSCs)and the CD106- subgroup.Methods The MSCs from normal BM were isolated and expanded.The subgroups of CD106+ and CD106-MSCs were sorted.The cell proliferation and adhesion functions,chemotactic activities,adipogenic and osteogenic potentials,senescence,and senescence protein 21(p21)were detected.The capacity of translocation into nucleus of nuclear factor-kappa B(NF-κB)when stimulated by tumor necrosis factor(TNF-α)was measured.Results The proliferative ability was higher in CD106+MSCs than that in CD106-MSCs.In 48 hours,the value of optical density(OD)was significantly higher in CD106+MSCs than that in CD106- subgroup(1.004±0.028 vs. 0.659±0.023,t=3.946,P=0.0225).In 72 hours,this phenomenon was even more pronounced(2.574±0.089 vs. 1.590±0.074,t=11.240,P=0.0000).The adhesive capacity of CD106+MSCs was significantly stronger than that of CD106- subgroup(0.648±0.018 vs. 0.418±0.023,t=7.869,P=0.0002).Besides,the metastasis ability of CD106+MSCs were significantly stronger than that of CD106- subgroup(114.500±4.481 vs.71.000±4.435,t=6.900,P=0.0005).The CD106+MSCs had signifcnatly lower proportions of senescent cells.The expression of aging protein p21 in CD106+MSCs was significantly lower than that in CD106-MSCs [(17.560±1.421)% vs.(45.800±2.569)%,t=9.618,P=0.0000].Furthermore,there were no visible pigmenting cells after β-galactosidase staining in CD106+MSCs subgroup.However,in CD106-MSCs,some colored green cells were detected.The rate of NF-κB translocation into nucleus after stimulated by TNF-α was significantly higher in CD106+MSCs than CD106- MSCs [(37.780±3.268)% vs.(7.30±1.25)%,t=8.713,P=0.0001].Conclusion Bone marrow-derived CD106+MSCs possess more powerful biological functions than CD106-MSCs.

关 键 词：CD106^+间充质干细胞 增殖功能 黏附功能 趋化功能 衰老 

分 类 号：R318.0[医药卫生—生物医学工程]