本文選題：腎脂肪囊 + 脂肪間充質(zhì)干細(xì)胞　； 參考：《遼寧醫(yī)學(xué)院》2012年碩士論文

【摘要】：目的 探討熒光染料氯甲基苯甲酰氨(chlormethyl-benzamidodialkylcarbocyanine,CM-DiI)對(duì)大鼠腎脂肪囊來(lái)源脂肪間充質(zhì)干細(xì)胞(Kidney fat capsule adipose-derived mesenchymal stem cells,KFC-ADSCs)的標(biāo)記效果，并分析評(píng)價(jià)該標(biāo)記方法對(duì)KFC-ADSCs的細(xì)胞形態(tài)、生長(zhǎng)及增殖能力等生物學(xué)特性的影響，旨在為KFC-ADSCs移植的體內(nèi)示蹤與歸巢研究尋找一種實(shí)用而有效的細(xì)胞標(biāo)記方法。 方法 采用膠原酶消化法分離、培養(yǎng)SD大鼠KFC-ADSCs；流式細(xì)胞儀檢測(cè)KFC-ADSCs表面標(biāo)記物；無(wú)菌、避光條件下采用CM-DiI細(xì)胞標(biāo)記液標(biāo)記大鼠第3代KFC-ADSCs，倒置熒光顯微鏡下觀察、計(jì)算細(xì)胞的標(biāo)記率及標(biāo)記的持續(xù)時(shí)間；MTT法分析比較CM-DiI標(biāo)記液對(duì)KFC-ADSCs的細(xì)胞形態(tài)、生長(zhǎng)及增殖等生物學(xué)特性是否存在影響。 結(jié)果 流式細(xì)胞儀檢測(cè)KFC-ADSCs細(xì)胞表面標(biāo)記：間充質(zhì)干細(xì)胞表面相關(guān)抗原CD29、CD44呈強(qiáng)陽(yáng)性表達(dá)，內(nèi)皮細(xì)胞表面相關(guān)抗原CD31呈陰性表達(dá)。倒置熒光顯微鏡下觀察CM-DiI標(biāo)記的KFC-ADSCs細(xì)胞膜及細(xì)胞漿被染成紅色，細(xì)胞核不著色。觀察CM-DiI標(biāo)記的KFC-ADSCs細(xì)胞標(biāo)記率達(dá)95%以上，，比較KFC-ADSCs標(biāo)記前后細(xì)胞的形態(tài)、生長(zhǎng)及增殖等生物學(xué)特性無(wú)明顯改變。CM-DiI標(biāo)記的KFC-ADSCs經(jīng)多次傳代培養(yǎng)后，紅色熒光細(xì)胞數(shù)量及強(qiáng)度有所下降，但細(xì)胞傳代至第8代、細(xì)胞標(biāo)記的第15d，CM-DiI細(xì)胞標(biāo)記率仍可達(dá)50%以上。 結(jié)論 CM-DiI可以有效、穩(wěn)定的標(biāo)記KFC-ADSCs，標(biāo)記方法簡(jiǎn)單，標(biāo)記效率高，對(duì)KFC-ADSCs無(wú)毒性作用，適用于KFC-ADSCs的中、短期標(biāo)記及KFC-ADSCs移植的體內(nèi)示蹤觀察。
[Abstract]:Purpose To investigate the labeling effect of fluorescent dye chlormethyl-benzamidalkylcarbocyanine (CM-DiI) on adipose mesenchymal stem cells (KFC-ADSCs) derived from renal fat sac in rats, and to analyze and evaluate the cell morphology of KFC-ADSCs by this method. The purpose of this study is to find a practical and effective method for the study of KFC-ADSCs transplantation in vivo for tracing and homing. Method KFC-ADSCs of SD rats were isolated by collagenase digestion, KFC-ADSCs surface markers were detected by flow cytometry, KFC-ADSCswere labeled with CM-DiI cell labeled solution in the condition of sterility, and observed under inverted fluorescence microscope. The cell labeling rate and the labeling duration were calculated. The effects of CM-DiI labeling solution on the cell morphology, growth and proliferation of KFC-ADSCs were compared. Result The surface markers of KFC-ADSCs cells were detected by flow cytometry: CD29 CD44 was strongly expressed on mesenchymal stem cells and CD31 was negative on endothelial cells. The CM-DiI labeled KFC-ADSCs cell membrane and cytoplasm were stained red and the nucleus was not stained under inverted fluorescence microscope. The labeling rate of KFC-ADSCs cells labeled with CM-DiI was more than 95%. The number and intensity of red fluorescent cells decreased after repeated passage culture of KFC-ADSCs labeled with KFC-ADSCs. However, the labeling rate of CM-DiI cells was still over 50% on the 15th day after passage to the 8th passage. Conclusion CM-DiI can effectively and stably label KFC-ADSCs.The labeling method is simple, the labeling efficiency is high, and it has no toxic effect on KFC-ADSCs. It is suitable for the in vivo tracer observation of middle and short term labeling of KFC-ADSCs and KFC-ADSCs transplantation.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R329

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