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作 者:吴绍宏[1] 甘建和[1] 秦爱兰[1] 黄小平[1] 罗二平[1] 赵卫峰[1] 江敏华[1]
机构地区:[1]苏州大学附属第一医院感染科,江苏苏州215006
出 处:《苏州大学学报(医学版)》2010年第5期918-921,F0003,共5页Suzhou University Journal of Medical Science
基 金:科技部"艾滋病和病毒性肝炎等重大传染病防治"科技重大专项"十一五"(2008ZX10002-005);苏州市科技计划项目(SS0704)
摘 要:目的分离、培养大鼠骨髓间充质干细胞(MSCs),研究肿瘤坏死因子(TNF-α)等因素对细胞表面趋化因子受体-4(CXCR4)表达的影响。方法全骨髓贴壁筛选法培养大鼠MSCs,成脂诱导鉴定;采用TNF-α等因子干预培养,流式细胞术检测胞膜CXCR4的表达。结果 MSCs呈梭形、漩涡状方式贴壁生长,成脂诱导后油红O染色可见胞浆内存在大量脂滴。肝衰竭大鼠血浆及TNF-α、白细胞介素-6(IL-6)、脂多糖(LPS)干预培养后MSCs胞膜的CXCR4表达率分别为(59.46±1.01)%、(56.34±0.89)%、(51.34±0.85)%和(44.66±0.36)%,与正常培养组的(26.80±0.35)%及大鼠血浆干预组的(27.13±0.67)%比较,差异均有统计学意义(均P<0.05)。结论全骨髓贴壁筛选法能够分离培养出纯度及活性较高的MSCs;TNF-α、IL-6等因素可以诱导MSCs胞膜CXCR4的表达。Objective To separate and cultivate the MSCs from rat bone marrow,and to study the influence of some intervention factors to the expression of CXCR4 on the surface of MSCs. Methods MSCs were isolated from rat bone marrow and cultured with cell culture technique,then adipogenic differentiation identification was carried out. MSCs were stimulated with IL-6,LPS or TNF-alpha respectively. Cell-surface expression of CXCR4 was assessed by flow cytometry. Results Rat MSCs looks fusiform under light microscope. The expression of CXCR4 of MSCs in the groups in which stimulation with plasma from acute liver failure,TNF-α,IL-6 and LPS was ( 59. 46 ± 1. 01) % ,( 56. 34 ± 0. 89) % ,( 51. 34 ± 0. 85) % and ( 44. 66 ± 0. 36) % ,respectively. While in common culture medium,the expression of CXCR4 is ( 26. 80 ± 0. 35) % ,which had statistical difference between the four stimulated groups and the control group ( P 0. 05) . Otherwise,those intervention factors also affected the proliferation of MSCs. Conclusions Rat MSCs can be succesfully isolated and cultured by whole bone marrow adherent culture. Stimulating factors such as TNF-α can enhance the expression of CXCR4 on the surface of MSCs.
关 键 词:大鼠 骨髓间充质干细胞 趋化因子受体-4 白细胞介素-6 肿瘤坏死因子-α 内毒素 流式细胞术 肝衰竭
分 类 号:R329.28[医药卫生—人体解剖和组织胚胎学]
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