银屑病患者DMSCs诱导人永生化角质形成细胞株的异常增殖

发布时间：2018-05-04 00:31

  本文选题：银屑病 + 人DMSCs细胞　； 参考：《山西医科大学》2015年硕士论文

【摘要】：目的:利用Transwell小室技术共同培养人真皮间充质干细胞(Human dermis mesenchymal stem cells,DMSCs)与Ha-cat细胞(人永生化角质形成细胞株);探明人DMSCs对Ha-cat细胞增殖的作用。方法:1、机械-酶顺序分离法分离银屑病患者和正常人的真皮与表皮,培养、纯化DMSCs;2、采用FCM检测Ha-cat细胞表面标记物;3、多向分化法诱导人DMSCs细胞分化,鉴定细胞的生物学特性;4、采用RTCA(实时动态细胞分析系统)检测Ha-cat细胞的增殖曲线;5、检测DMSCs诱导后的Ha-cat细胞的活细胞数量;6、用FCM检测Ha-cat细胞的增殖周期。结果:1、银屑病患者组与正常人组DMSCs细胞在形态与培养方法上无显著的差异;2、FCM检测人DMSCs表面标记物:CD29、CD44、CD73、CD90、CD105及CD105均呈高表达,而CD34、CD45及HLA-DR表达阴性,表明DMSCs有较高的纯度;3、成功使用化学试剂诱导DMSCs定向分化,使之分化为脂肪等多种组织细胞;4、x CELLigence检测结果表明:银屑病患者组较正常人对照组DMSCs细胞促进Ha-cat细胞的增殖;(患者组Ha-cat细胞CIMax值为5.55,对照组Ha-cat细胞CIMax值为5.40)5、细胞计数仪检测:银屑病患者组Ha-cat活细胞数较正常对照组有所增加;6、FCM检测细胞的增殖周期:银屑病组Ha-cat细胞进入S期的细胞比例增加。(p0.05)。结论:1、成功模拟生物状态下人DMSCs与Ha-cat细胞(人永生化角质形成细胞株)的相互作用;2、银屑病表皮的异常增厚与人DMSCs相关。
[Abstract]:Aim: to investigate the effect of human DMSCs on the proliferation of human dermal mesenchymal stem cells (DMSCs) and Ha-cat cells (human immortalized keratinocytes) by co-culture of human dermal mesenchymal stem cells (dermis mesenchymal stem cells) with Transwell chamber technique. Methods the dermis and epidermis of psoriasis patients and normal persons were separated by mechanical-enzymatic sequential separation method. DMSCs2 was cultured and purified. The surface marker of Ha-cat cells was detected by FCM. The differentiation of human DMSCs cells was induced by multiple differentiation method. The biological characteristics of the cells were identified. The proliferation curve of Ha-cat cells was detected by RTCA (Real time dynamic Cell Analysis system), the number of living cells induced by DMSCs was detected and the proliferative cycle of Ha-cat cells was detected by FCM. Results there was no significant difference in morphology and culture methods of DMSCs cells between psoriatic patients and normal controls. FCM was used to detect the high expression of CD105 and CD105 in human DMSCs surface markers: 1. The expression of CD34 + CD45 and HLA-DR was negative. The results showed that DMSCs had a high purity and was successfully induced by chemical reagents to induce the directional differentiation of DMSCs. The results of CELLigence analysis showed that DMSCs cells in psoriatic patients promoted the proliferation of Ha-cat cells (CIMax value of Ha-cat cells was 5.55, CIMax value of Ha-cat cells in control group was 5.40%, and that of control group was 5.40%). The number of Ha-cat living cells in psoriatic patients was increased compared with the control group. The proliferative cycle of Ha-cat cells in psoriasis group was increased. The percentage of Ha-cat cells entering S phase was increased in psoriasis group. Conclusion the interaction between human DMSCs and Ha-cat cells (human immortalized keratinocyte line) was successfully simulated under biological condition. The abnormal thickening of psoriatic epidermis was related to human DMSCs.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R758.63

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