【摘要】：目的:探尋小鼠ADSCs的體外分離培養(yǎng)方法以及成骨誘導(dǎo)前后造血調(diào)控因子分子水平的表達(dá)情況。方法:采用酶消化法獲得脂肪來(lái)源的細(xì)胞,傳代培養(yǎng)至第三代,分別進(jìn)行成脂與成骨細(xì)胞誘導(dǎo),用實(shí)時(shí)熒光定量多聚酶鏈反應(yīng)(qRT-PCR)檢測(cè)成骨誘導(dǎo)前后的造血調(diào)控因子β-catenin、SDF-1、N-cadherin、SCF、MIP-1α和Jag 1,GM-CSF的表達(dá)。同時(shí)采用流式細(xì)胞儀測(cè)定ADSCs細(xì)胞表面標(biāo)志物CD29、CD34、CD106、CD49d的表達(dá)。結(jié)果:小鼠脂肪來(lái)源的細(xì)胞在體外培養(yǎng)過(guò)程中,呈梭形貼壁生長(zhǎng),可穩(wěn)定傳代;細(xì)胞表面標(biāo)志的流式細(xì)胞儀檢測(cè):CD29表達(dá)率強(qiáng)陽(yáng)性,CD34表達(dá)率較低,CD106、CD49d低表達(dá);進(jìn)行成脂細(xì)胞誘導(dǎo)后,油紅“O”染色呈陽(yáng)性,成骨細(xì)胞誘導(dǎo)后,茜素紅染色和von Kossa染色均為陽(yáng)性,表明其是ADSCs。通過(guò)qRT-PCR檢測(cè)多種造血調(diào)控因子相對(duì)表達(dá)量,發(fā)現(xiàn)β-catenin表達(dá)量最高,SDF-1和N-cadherin次之,其余各因子SCF、MIP-1α、Jag1、GM-CSF均為低表達(dá)。統(tǒng)計(jì)分析后,發(fā)現(xiàn)各因子在成骨誘導(dǎo)前后表達(dá)差異均無(wú)統(tǒng)計(jì)學(xué)意義。結(jié)論:本研究證實(shí),小鼠脂肪來(lái)源的細(xì)胞具有MSC特征就是ADSCs,其成骨誘導(dǎo)前后的造血調(diào)控因子均有不同程度的表達(dá),但差異無(wú)統(tǒng)計(jì)學(xué)意義。
[Abstract]:Aim: to investigate the isolation and culture of mouse ADSCs in vitro and the expression of hematopoietic regulatory factors before and after osteogenesis. Methods: the adipose derived cells were obtained by enzyme digestion and cultured to the third generation. The adipogenic cells and osteoblasts were induced respectively. The expression of hematopoietic regulatory factors 尾-catenin,SDF-1,N-cadherin,SCF,MIP-1 偽 and Jag 1 GM-CSF before and after osteogenesis was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). At the same time, flow cytometry was used to detect the expression of CD29,CD34,CD106,CD49d on the surface of ADSCs cells. Results: the murine adipose derived cells were fusiform adherent to the wall in vitro and could be subcultured stably. Flow cytometry showed that the expression rate of CD29 was strongly positive, the expression rate of CD34 was low, and the expression of CD106,CD49d was low. After adipoblast induction, oil red "O" staining was positive, and osteoblast induction, alizarin red staining and von Kossa staining were positive, indicating that it was ADSCs.. The relative expression of various hematopoietic regulatory factors was detected by qRT-PCR. It was found that the expression of 尾-catenin was the highest, followed by SDF-1 and N-cadherin, and the other factors SCF,MIP-1 偽 and Jag1,GM-CSF were all low expression. After statistical analysis, it was found that there was no significant difference in the expression of each factor before and after osteogenesis. Conclusion: this study confirmed that the expression of hematopoietic regulatory factors before and after osteogenesis induced by ADSCs, was the characteristic of MSC in adipose derived cells of mice, but the difference was not statistically significant.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周蘇娜;張明鑫;王健生;;脂肪來(lái)源的間充質(zhì)干細(xì)胞的生物學(xué)特征及臨床應(yīng)用[J];中國(guó)現(xiàn)代普通外科進(jìn)展;2009年01期

2 張春華;溫澤清;朱勇;陳秀玲;;人骨髓造血干細(xì)胞體外擴(kuò)增培養(yǎng)及其生物學(xué)特性研究[J];山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2006年10期

3 石小玲;;集落刺激因子對(duì)造血的調(diào)控作用及其檢測(cè)的臨床意義[J];實(shí)用醫(yī)技雜志;2007年15期

4 張龔莉;李寧;宋永平;林全德;魏旭東;房佰俊;;脂肪源性間充質(zhì)干細(xì)胞聯(lián)合造血干細(xì)胞共移植治療頑固性重型再生障礙性貧血2例(英文)[J];中國(guó)組織工程研究與臨床康復(fù);2009年36期

5 丁志;楊松林;;間充質(zhì)干細(xì)胞生物學(xué)特性及其分化潛能[J];中國(guó)組織工程研究與臨床康復(fù);2011年01期

6 畢曉娟;馬艷;江明;;大鼠腹股溝脂肪間充質(zhì)干細(xì)胞的分離、培養(yǎng)、鑒定及活體標(biāo)記[J];中國(guó)組織工程研究與臨床康復(fù);2011年14期

7 唐佩弦;再論造血干細(xì)胞研究的若干新動(dòng)向[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2001年01期

8 黃曉兵;劉霆;孟文彤;智偉;;骨髓間充質(zhì)干細(xì)胞分化的成骨細(xì)胞支持臍血造血干祖細(xì)胞的研究[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2006年03期

9 于立志，，袁長(zhǎng)吉;造血干細(xì)胞模仿體內(nèi)擴(kuò)增研究進(jìn)展[J];中華血液學(xué)雜志;1995年11期

10 崔磊,尹爍,楊平,劉波,張英,劉偉,曹誼林;脂肪干細(xì)胞HLA分子表達(dá)與體外抑制淋巴細(xì)胞增殖的實(shí)驗(yàn)研究[J];中華醫(yī)學(xué)雜志;2005年27期

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