本文選題：外胚間充質(zhì)細(xì)胞 + 成骨細(xì)胞　； 參考：《昆明醫(yī)學(xué)院》2008年碩士論文

【摘要】： 目的:建立SD胎鼠頜突未分化外胚間充質(zhì)細(xì)胞的體外培養(yǎng)細(xì)胞模型,在此基礎(chǔ)上探討胰島素樣生長因子-1(IGF-1)誘導(dǎo)胎鼠頜突外胚間充質(zhì)細(xì)胞向成骨細(xì)胞(OB)分化的作用。 方法:采用消化貼壁法培養(yǎng)SD胎鼠外胚間充質(zhì)細(xì)胞,建立外胚間充質(zhì)細(xì)胞的體外培養(yǎng)細(xì)胞模型。將生長狀態(tài)良好的第三代頜突間充質(zhì)細(xì)胞培養(yǎng)于含有20 ng/mlIGF-1、50 ml/L胎牛血清(FBS)的DMEM/F12培養(yǎng)基中,連續(xù)培養(yǎng)10d。用相差倒置顯微鏡觀察細(xì)胞的生長和形態(tài)變化,HE染色,觀察細(xì)胞形態(tài)變化。抗I型膠原單抗免疫組化染色鑒定細(xì)胞性質(zhì),堿性磷酸酶染色及活性檢測。 結(jié)果:原代培養(yǎng)的間充質(zhì)細(xì)胞呈纖維樣細(xì)胞生長,免疫組化染色,抗VIM為陽性,抗CK為陰性。經(jīng)IGF-1培養(yǎng)的外胚間充質(zhì)細(xì)胞,10d后在倒置顯微鏡下可見細(xì)胞形態(tài)發(fā)生明顯變化,呈多邊形,細(xì)胞緊密排列,伸出多個(gè)突起,似成骨細(xì)胞。細(xì)胞爬片HE染色,可見細(xì)胞呈不規(guī)則形態(tài),邊緣伸出多個(gè)突起�？笽型膠原單抗(Collage-I)免疫組化染色陽性,堿性磷酸酶(ALP)染色陽性,堿性磷酸酶(ALP)活性增高。 結(jié)論:頜突外胚間充質(zhì)細(xì)胞經(jīng)胰島素樣生長因子-1(IGF-1)的誘導(dǎo)培養(yǎng),細(xì)胞形態(tài)出現(xiàn)類成骨樣改變,同時(shí)細(xì)胞分泌堿性磷酸酶和I型膠原,出現(xiàn)類似成骨細(xì)胞的功能性改變。
[Abstract]:Aim: to establish an in vitro culture model of undifferentiated extracellular mesenchymal cells from maxillary process of SD fetal rats, and to investigate the effect of insulin-like growth factor-1 (IGF-1) on differentiation of mesenchymal cells from maxillary process to osteoblasts (OB). Methods: the mesenchymal cells of SD rat embryo were cultured by digestion and adherent method, and the model of cultured cells was established in vitro. The third generation of maxillary process mesenchymal cells were cultured in DMEM / F12 medium containing 20 ng / ml IGF-1 / 50 ml / L fetal bovine serum (FBS) for 10 days. Cell growth and morphological changes were observed by phase contrast inverted microscope. Immunohistochemical staining of anti-type I collagen monoclonal antibody was used to identify cell properties, alkaline phosphatase staining and activity detection. Results: the primary cultured mesenchymal cells grew as fibroid cells, and immunohistochemical staining showed that anti-vim was positive and anti-CK was negative. After 10 days of IGF-1 culture, obvious changes of cell morphology were observed under inverted microscope, with polygon, tight arrangement of cells, multiple protrusions and osteoblast-like cells. Cell climbing slice HE staining, the cells showed irregular shape, the edge of multiple protrusions. The immunohistochemical staining of anti type I collagen monoclonal antibody (Collage-I) and alkaline phosphatase (ALP) were positive, and the activity of alkaline phosphatase (ALP) was increased. Conclusion: induced by insulin-like growth factor-1 (IGF-1), osteoblast-like changes were observed in the mesenchymal cells of the maxillary process, and alkaline phosphatase and collagen I were secreted by the cells, and the functional changes were similar to those of the osteoblasts.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前9條

1 杜俊杰,羅卓荊,胡蘊(yùn)玉,李志創(chuàng),呂榮;rhBMP-2在體內(nèi)誘導(dǎo)成骨中I,II型膠原及堿性磷酸酶的表達(dá)[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2001年11期

2 王有偉,陳以慈;堿性磷酸酶染色法(鈣鉛法)在研究人腦和心臟微血管方面的應(yīng)用[J];解剖學(xué)雜志;1986年03期

3 呂紅兵,金巖,李媛,趙宇;BALB/c胎鼠面突未分化外胚間充質(zhì)細(xì)胞的體外培養(yǎng)[J];實(shí)用口腔醫(yī)學(xué)雜志;1999年01期

4 吳拓江;應(yīng)力作用下骨縫中成骨細(xì)胞的細(xì)胞因子改變[J];現(xiàn)代口腔醫(yī)學(xué)雜志;2004年01期

5 陸懷秀,趙宇,肖明振,牛忠英,郭紅延;大鼠上下頜突未分化外胚間充質(zhì)細(xì)胞的體外培養(yǎng)[J];牙體牙髓牙周病學(xué)雜志;2001年03期

6 鄧蔓菁,金巖,史俊南,呂紅兵,劉源,趙宇;酶消化法培養(yǎng)Balb/c胎鼠下頜突未分化外胚間充質(zhì)細(xì)胞[J];牙體牙髓牙周病學(xué)雜志;2002年08期

7 鄧蔓菁,金巖,史俊南,呂紅兵,劉源,趙宇;Balb/c胎鼠下頜突外胚間充質(zhì)細(xì)胞向成骨細(xì)胞誘導(dǎo)分化的體外研究[J];牙體牙髓牙周病學(xué)雜志;2003年05期

8 曾融生;束煌;王劍寧;;rhBMP-2誘導(dǎo)成肌細(xì)胞表達(dá)成骨表型的研究[J];中國口腔頜面外科雜志;2007年01期

9 劉大為,傅民魁,李盛琳,章魁華,吳燕婉;機(jī)械牽張作用對(duì)UMR-106細(xì)胞骨橋蛋白mRNA和TGF-β1 mRNA表達(dá)的影響[J];中華口腔醫(yī)學(xué)雜志;2000年01期

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