本文關(guān)鍵詞： 骨髓間充質(zhì)干細(xì)胞 細(xì)胞培養(yǎng) Ⅰ型膠原蛋白 成骨分化 兔　出處：《青島大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的通過(guò)實(shí)驗(yàn)進(jìn)一步探討體外條件下分離、純化兔骨髓間充質(zhì)干細(xì)胞的有效方法,對(duì)所獲細(xì)胞進(jìn)行培養(yǎng),觀察細(xì)胞的生物學(xué)特性。以及觀察Ⅰ型膠原蛋白對(duì)兔骨髓間充質(zhì)干細(xì)胞成骨分化的作用。 方法實(shí)驗(yàn)于青島大學(xué)醫(yī)學(xué)院附屬醫(yī)院骨科研究所進(jìn)行。首先采用密度梯度離心法分離兔骨髓間充質(zhì)干細(xì)胞,用貼壁培養(yǎng)法對(duì)所獲細(xì)胞進(jìn)行培養(yǎng)、傳代、純化和擴(kuò)增,觀察細(xì)胞的生物學(xué)特性,倒置相差顯微鏡觀察細(xì)胞的形態(tài)學(xué)特征,透射電鏡觀察細(xì)胞的超微結(jié)構(gòu),流式細(xì)胞術(shù)鑒定細(xì)胞表面標(biāo)志物;將分離純化的兔骨髓間充質(zhì)干細(xì)胞培養(yǎng)于Ⅰ型膠原蛋白上面,對(duì)細(xì)胞的堿性磷酸酶表達(dá)、細(xì)胞礦化形成鈣化結(jié)節(jié)的能力進(jìn)行測(cè)定。 結(jié)果分離的細(xì)胞1天后完全貼壁,貼壁的細(xì)胞平均10d后形成細(xì)胞克隆,所獲細(xì)胞形態(tài)均一,呈成纖維細(xì)胞樣生長(zhǎng),細(xì)胞穩(wěn)定表達(dá)骨髓間充質(zhì)干細(xì)胞表面標(biāo)志物。在Ⅰ型膠原蛋白上面培養(yǎng)后,細(xì)胞堿性磷酸酶顯著表達(dá),Von.Kossa染色可見(jiàn)礦化結(jié)節(jié)形成。 結(jié)論密度梯度離心法分離,貼壁培養(yǎng)法培養(yǎng)所得細(xì)胞,可以有效分離、純化兔骨髓間充質(zhì)干細(xì)胞。用此種方法獲得的細(xì)胞具有骨髓間充質(zhì)干細(xì)胞的特性。Ⅰ型膠原蛋白能夠促進(jìn)兔骨髓間充質(zhì)干細(xì)胞的成骨分化。
[Abstract]:Objective to investigate the effective method of isolation and purification of rabbit bone marrow mesenchymal stem cells (BMSCs) in vitro. To observe the biological characteristics of the cells and the effect of type I collagen on the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells. Methods the experiment was carried out in the Institute of Orthopaedics, affiliated Hospital of Qingdao University Medical College. Firstly, bone marrow mesenchymal stem cells were isolated by density gradient centrifugation. The biological characteristics of cells were observed by purification and amplification, the morphological characteristics of cells were observed by inverted phase contrast microscope, the ultrastructure of cells was observed by transmission electron microscopy, and cell surface markers were identified by flow cytometry. The purified rabbit bone marrow mesenchymal stem cells were cultured on type I collagen. The expression of alkaline phosphatase and the ability of mineralization to form calcified nodules were measured. Results the isolated cells were completely adhered to the wall one day later, and the adherent cells formed cell clones after an average of 10 days. The obtained cells were homogeneous in morphology and grew like fibroblasts. The surface markers of bone marrow mesenchymal stem cells were stably expressed in the cells. After cultured on type 鈪，

本文編號(hào)：1467130