【摘要】：目的：探討在體外條件下不同濃度的利福噴丁溶液對兔脂肪干細(xì)胞增殖活性的影響。方法：本試驗(yàn)取健康新西蘭大白兔(雌雄不限)，體重2.5-3.0kg，由新疆醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心提供。用3％戊巴比妥鈉(1ml/kg)或速眠新(0.3ml/kg)麻醉后，8％硫化鈉水溶液術(shù)區(qū)脫毛備皮，消毒，取肩胛間區(qū)皮下脂肪，無菌切取皮下脂肪。放入裝有10ml PBS的培養(yǎng)皿中，帶回實(shí)驗(yàn)室。用I型膠原酶將細(xì)胞消化后，接種在DMEM生長液中進(jìn)行培養(yǎng)。在倒置顯微鏡下觀察細(xì)胞生長態(tài)勢以及形狀。待細(xì)胞增殖到一定程度時(shí)進(jìn)行傳代，取第三代細(xì)胞接種后加入成骨誘導(dǎo)劑誘導(dǎo)成骨，于成骨誘導(dǎo)第17天鏡下觀察有鈣結(jié)節(jié)形成，茜素紅染色結(jié)果陽性，，證實(shí)所取細(xì)胞為兔脂肪干細(xì)胞，然后再取第三代兔脂肪干細(xì)胞分別設(shè)3組不同利福噴丁濃度(根據(jù)每組利福噴丁濃度不同，分為10μg/ml組、20μg/ml組、40μg/ml組)及空白對照組進(jìn)行細(xì)胞培養(yǎng)，采用CCK-8比色法進(jìn)行增殖分化活性比較。用堿性磷酸酶及礦化結(jié)節(jié)染色鑒定成骨細(xì)胞分化能力。結(jié)果：堿性磷酸酶及礦化結(jié)節(jié)染色均為陽性。與空白對照組相比：利福噴丁溶液濃度為10μg/ml、20μg/ml、40μg/ml時(shí)細(xì)胞增殖活性在統(tǒng)計(jì)學(xué)上無差異(P＞0.05)；在顯微鏡下可觀察到利福噴丁溶液濃度為40μg/ml時(shí)兔脂肪干細(xì)胞形態(tài)發(fā)生明顯變化。結(jié)論：從兔脂肪組織中分離、培養(yǎng)出的脂肪干細(xì)胞(ADSCs)在具有在體外誘導(dǎo)成為成骨細(xì)胞(OB)的分化潛能。利福噴丁溶液濃度為10μg/ml、20μg/ml、40μg/ml時(shí)細(xì)胞增殖活性在統(tǒng)計(jì)學(xué)上無差異(P＞0.05)；在顯微鏡下可觀察到利福噴丁溶液濃度為40μg/ml時(shí)兔脂肪干細(xì)胞形態(tài)發(fā)生明顯變化。
[Abstract]:Aim: to investigate the effect of rifapentine solution at different concentrations on the proliferation of rabbit adipose stem cells in vitro. Methods: the healthy New Zealand white rabbits (male and female), weighing 2.5-3.0 kg, were provided by Experimental Animal Center of Xinjiang Medical University. After anaesthesia with 3% pentobarbital sodium (1ml/kg) or 0.3ml/kg, 8% sodium sulphide solution was used to remove hair and skin, sterilize, take subcutaneous fat in interscapular area, and cut subcutaneous fat aseptically. Put it in a petri dish containing 10ml PBS and bring it back to the lab. Cells were digested with type I collagenase and cultured in DMEM growth liquid. Cell growth and shape were observed under inverted microscope. After the third generation cells were inoculated with osteogenic inducer to induce osteogenesis, calcium nodules were observed under the microscope on the 17th day of osteogenesis induction, and alizarin red staining was positive. The cells were confirmed to be rabbit adipose stem cells, and then the third generation of rabbit adipose stem cells were divided into three groups with different concentrations of rifapentine (10 渭 g/ml group, 20 渭 g/ml group, 40 渭 g/ml group) and blank control group, respectively. The activity of proliferation and differentiation was compared by CCK-8 colorimetry. The differentiation ability of osteoblasts was evaluated by alkaline phosphatase and mineralized nodule staining. Results: alkaline phosphatase and mineralized nodules were positive. When the concentration of rifapentine solution was 10 渭 g / ml or 20 渭 g / ml or 40 渭 g/ml, the proliferative activity of rabbit adipose stem cells had no statistical difference (P > 0. 05), and the morphological changes of adipose stem cells were observed under microscope when the concentration of rifapentine solution was 40 渭 g/ml. Conclusion: adipose stem cell (ADSCs) isolated from rabbit adipose tissue has the potential to differentiate into osteoblast (OB) in vitro. When the concentration of rifapentine solution was 10 渭 g / ml ~ (20 渭 g / ml), the proliferative activity of rabbit adipose stem cells had no statistical difference (P > 0. 05), and the morphological changes of adipose stem cells were observed under microscope when the concentration of rifapentine solution was 40 渭 g/ml.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 翟廣;李波;李玉芹;劉亞男;寇長貴;姚燕;趙書欣;;利福噴丁與利福平治療肺結(jié)核療效和安全性的Meta分析[J];吉林大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2011年03期

2 郝偉;胡蘊(yùn)玉;姜明;呂榮;王軍;趙廷寶;;基于脂肪干細(xì)胞I型膠原凝膠PLGA-β-TCP支架的新型仿生骨組織工程復(fù)合體的構(gòu)建及生物學(xué)性能[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2008年07期

3 藍(lán)旭;文益民;葛寶豐;劉雪梅;;骨髓基質(zhì)干細(xì)胞復(fù)合納米羥基磷灰石/聚乳酸修復(fù)節(jié)段性骨缺損[J];中國骨與關(guān)節(jié)損傷雜志;2006年10期

4 夏萬堯;孫恒峗;陳瑾君;劉偉;周廣東;曹誼林;;HTGF-β1基因轉(zhuǎn)染脂肪干細(xì)胞及其表達(dá)[J];解剖科學(xué)進(jìn)展;2011年02期

5 王�？�;趙德萍;代曉明;李逸松;何曉光;劉流;;不同類型血清對大鼠脂肪干細(xì)胞分離培養(yǎng)的影響[J];昆明醫(yī)學(xué)院學(xué)報(bào);2010年03期

6 李相杰;;利福噴丁、利福平治療菌陽肺結(jié)核的療效觀察[J];臨床肺科雜志;2006年03期

7 高曉鳳;李健;李幼平;;利福布丁較利福噴丁治療耐多藥肺結(jié)核的系統(tǒng)評價(jià)[J];川北醫(yī)學(xué)院學(xué)報(bào);2009年03期

8 程新平;車立新;李坤;伊力哈爾;;新疆骨結(jié)核臨床特點(diǎn)初步分析[J];實(shí)用骨科雜志;2011年05期

9 蔣新華;陳建宇;蔡兆熙;張婭;劉珍珍;謝傳淼;伍堯泮;吳沛宏;;SPIO標(biāo)記脂肪干細(xì)胞移植退變椎間盤內(nèi)的MRI活體示蹤[J];中國組織工程研究與臨床康復(fù);2011年40期

10 刁琳鑒;;利福布汀用于耐多藥肺結(jié)核病臨床效果觀察[J];醫(yī)學(xué)信息(中旬刊);2011年05期

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