本文選題：銀屑病 + 人DMSCs細(xì)胞��； 參考：《山西醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:利用Transwell小室技術(shù)共同培養(yǎng)人真皮間充質(zhì)干細(xì)胞(Human dermis mesenchymal stem cells,DMSCs)與Ha-cat細(xì)胞(人永生化角質(zhì)形成細(xì)胞株);探明人DMSCs對(duì)Ha-cat細(xì)胞增殖的作用。方法:1、機(jī)械-酶順序分離法分離銀屑病患者和正常人的真皮與表皮,培養(yǎng)、純化DMSCs;2、采用FCM檢測(cè)Ha-cat細(xì)胞表面標(biāo)記物;3、多向分化法誘導(dǎo)人DMSCs細(xì)胞分化,鑒定細(xì)胞的生物學(xué)特性;4、采用RTCA(實(shí)時(shí)動(dòng)態(tài)細(xì)胞分析系統(tǒng))檢測(cè)Ha-cat細(xì)胞的增殖曲線;5、檢測(cè)DMSCs誘導(dǎo)后的Ha-cat細(xì)胞的活細(xì)胞數(shù)量;6、用FCM檢測(cè)Ha-cat細(xì)胞的增殖周期。結(jié)果:1、銀屑病患者組與正常人組DMSCs細(xì)胞在形態(tài)與培養(yǎng)方法上無(wú)顯著的差異;2、FCM檢測(cè)人DMSCs表面標(biāo)記物:CD29、CD44、CD73、CD90、CD105及CD105均呈高表達(dá),而CD34、CD45及HLA-DR表達(dá)陰性,表明DMSCs有較高的純度;3、成功使用化學(xué)試劑誘導(dǎo)DMSCs定向分化,使之分化為脂肪等多種組織細(xì)胞;4、x CELLigence檢測(cè)結(jié)果表明:銀屑病患者組較正常人對(duì)照組DMSCs細(xì)胞促進(jìn)Ha-cat細(xì)胞的增殖;(患者組Ha-cat細(xì)胞CIMax值為5.55,對(duì)照組Ha-cat細(xì)胞CIMax值為5.40)5、細(xì)胞計(jì)數(shù)儀檢測(cè):銀屑病患者組Ha-cat活細(xì)胞數(shù)較正常對(duì)照組有所增加;6、FCM檢測(cè)細(xì)胞的增殖周期:銀屑病組Ha-cat細(xì)胞進(jìn)入S期的細(xì)胞比例增加。(p0.05)。結(jié)論:1、成功模擬生物狀態(tài)下人DMSCs與Ha-cat細(xì)胞(人永生化角質(zhì)形成細(xì)胞株)的相互作用;2、銀屑病表皮的異常增厚與人DMSCs相關(guān)。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R758.63

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本文編號(hào)：1840799