本文關(guān)鍵詞： 脂肪組織 干細(xì)胞 傳代 誘導(dǎo)分化 骨組織 生物學(xué)特性　出處：《蘭州大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:觀察體外長期培養(yǎng)對成人脂肪間充質(zhì)干細(xì)胞(ADMSCs)的基本生物學(xué)性狀及成骨分化潛能的影響。 方法:從成人脂肪組織分離培養(yǎng)ADMSCs,體外傳代擴(kuò)增,流式細(xì)胞術(shù)鑒定細(xì)胞表型,倒置相差顯微鏡及透視電子顯微鏡觀察細(xì)胞形態(tài)及超微結(jié)構(gòu),MTT法繪制生長曲線,成骨誘導(dǎo),堿性磷酸酶(ALP)染色、Von Kossa染色及免疫細(xì)胞化學(xué)染色行成骨鑒定,分析光密度值。 結(jié)果:成人ADMSCs表達(dá)CD44,不表達(dá)CD34,體外長期培養(yǎng)其形態(tài)、生長方式及超微結(jié)構(gòu)無明顯變化,成骨誘導(dǎo)后變?yōu)闄E圓形、島狀分布,ALP、鈣化結(jié)節(jié)、骨鈣素(OC)及Ⅰ型膠原染色均呈陽性,但隨著傳代次數(shù)的增加,光密度值有所減低。 結(jié)論:成人ADMSCs經(jīng)體外長期培養(yǎng)其一般生物學(xué)性狀可保持穩(wěn)定,但成骨分化潛能有所下降。
[Abstract]:Aim: to observe the effects of long-term culture on the basic biological characteristics and osteogenic differentiation potential of adult adipose mesenchymal stem cells (ADMSCs). Methods: ADMSCs were isolated and cultured from adult adipose tissue. The phenotypes of the cells were identified by flow cytometry. The morphology and ultrastructure of the cells were observed by inverted phase contrast microscope and fluoroscopic electron microscope. MTT method was used to draw the growth curve, osteogenesis induction, alkaline phosphatase (ALP) staining, Von Kossa staining and immunocytochemical staining for osteogenesis identification and optical density analysis. Results: adult ADMSCs expressed CD44, but not CD34. The morphology, growth pattern and ultrastructure of adult ADMSCs were not changed after osteogenesis induction. ALP, calcified nodules, osteocalcin (OC) and type I collagen staining were positive, but the optical density decreased with the increase of passage times. Conclusion: the general biological characteristics of adult ADMSCs cultured for a long time can be kept stable, but the osteogenic differentiation potential of adult ADMSCs can be decreased.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R329

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