本文選題：腎脂肪囊 + 脂肪間充質(zhì)干細胞�。� 參考：《遼寧醫(yī)學院》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)對大鼠腎脂肪囊來源脂肪間充質(zhì)干細胞(Kidney fat capsule adipose-derived mesenchymal stem cells,KFC-ADSCs)的標記效果，并分析評價該標記方法對KFC-ADSCs的細胞形態(tài)、生長及增殖能力等生物學特性的影響，旨在為KFC-ADSCs移植的體內(nèi)示蹤與歸巢研究尋找一種實用而有效的細胞標記方法。 方法 采用膠原酶消化法分離、培養(yǎng)SD大鼠KFC-ADSCs；流式細胞儀檢測KFC-ADSCs表面標記物；無菌、避光條件下采用CM-DiI細胞標記液標記大鼠第3代KFC-ADSCs，倒置熒光顯微鏡下觀察、計算細胞的標記率及標記的持續(xù)時間；MTT法分析比較CM-DiI標記液對KFC-ADSCs的細胞形態(tài)、生長及增殖等生物學特性是否存在影響。 結果 流式細胞儀檢測KFC-ADSCs細胞表面標記：間充質(zhì)干細胞表面相關抗原CD29、CD44呈強陽性表達，內(nèi)皮細胞表面相關抗原CD31呈陰性表達。倒置熒光顯微鏡下觀察CM-DiI標記的KFC-ADSCs細胞膜及細胞漿被染成紅色，細胞核不著色。觀察CM-DiI標記的KFC-ADSCs細胞標記率達95%以上，，比較KFC-ADSCs標記前后細胞的形態(tài)、生長及增殖等生物學特性無明顯改變。CM-DiI標記的KFC-ADSCs經(jīng)多次傳代培養(yǎng)后，紅色熒光細胞數(shù)量及強度有所下降，但細胞傳代至第8代、細胞標記的第15d，CM-DiI細胞標記率仍可達50%以上。 結論 CM-DiI可以有效、穩(wěn)定的標記KFC-ADSCs，標記方法簡單，標記效率高，對KFC-ADSCs無毒性作用，適用于KFC-ADSCs的中、短期標記及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.
【學位授予單位】：遼寧醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R329

【參考文獻】

相關期刊論文 前4條

1 馬慧雨;肖苒;杜曉巖;;熒光活性染料(CM-Dil)對人骨髓間充質(zhì)干細胞增殖能力的影響[J];口腔頜面外科雜志;2011年04期

2 吳畏;鄭紅光;張德偉;梅煜明;;腎脂肪囊來源脂肪干細胞的體外培養(yǎng)及其生物學特性的研究[J];細胞與分子免疫學雜志;2010年11期

3 鞠剛;徐衛(wèi)袁;張亞;張興祥;嚴飛;沙衛(wèi)平;;多孔絲素蛋白/羥基磷灰石復合脂肪間充質(zhì)干細胞修復兔關節(jié)軟骨及軟骨下骨缺損[J];中國組織工程研究與臨床康復;2011年29期

4 伍耀豪;劉廣鵬;劉波;崔磊;;應用自體脂肪干細胞修復豬關節(jié)軟骨缺損的初步實驗研究[J];組織工程與重建外科雜志;2006年03期

本文編號：1969512