本文關(guān)鍵詞： 肝干細(xì)胞 細(xì)胞分離 純化 免疫細(xì)胞化學(xué) 鑒定　出處：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2013年04期 　論文類(lèi)型：期刊論文

【摘要】：目的改進(jìn)大鼠胎肝干細(xì)胞的分離純化方法,提高肝臟干細(xì)胞分離的純度和活力。方法采用剪碎酶消化法分離大鼠胎肝干細(xì)胞,差速離心結(jié)合選擇性消化法進(jìn)行進(jìn)一步純化,含10%優(yōu)等胎牛血清的DMEM/F12培養(yǎng)液培養(yǎng)。利用免疫細(xì)胞化學(xué)方法分別檢測(cè)P1和P10代肝干細(xì)胞表面標(biāo)志物。結(jié)果分離的大鼠胎肝干細(xì)胞接種培養(yǎng)瓶24 h后開(kāi)始貼壁,5 d后細(xì)胞體積增大,呈梭形或多邊形。經(jīng)1～2次差異性消化傳代后,細(xì)胞呈集落樣生長(zhǎng),邊緣清晰且折光率強(qiáng)。免疫細(xì)胞化學(xué)檢測(cè)示:P1代大鼠胎肝干細(xì)胞C-Kit、甲胎蛋白(AFP)抗原、OV6、細(xì)胞角蛋白(CK)19呈陽(yáng)性表達(dá),不表達(dá)Alb。P10代肝干細(xì)胞C-Kit、CK19、OV-6及CD34呈陽(yáng)性表達(dá)。結(jié)論大鼠胎肝干細(xì)胞分離純化、培養(yǎng)擴(kuò)增及鑒定方法簡(jiǎn)單易行。
[Abstract]:Objective to improve the method of isolation and purification of rat fetal liver stem cells and improve the purity and activity of liver stem cells isolation. Methods Rat fetal liver stem cells were isolated by shearing enzyme digestion. Differential centrifugation combined with selective digestion for further purification. DMEM/F12 medium containing 10% superior fetal bovine serum was cultured. The surface markers of P1 and P10 passage liver stem cells were detected by immunocytochemistry. Culture bottle 24. H then began to adhere to the wall. After 5 days, the cells increased in volume and appeared fusiform or polygon. After two different digestion and passage, the cells grew like colony. The results of immunocytochemistry showed that C-Kitt, AFP) antigen OV6 and cytokeratin CK-19 were positive. There was no positive expression of C-Kit19 CK19 OV-6 and CD34 in Alb.P10 generation liver stem cells. Conclusion the methods of isolation, amplification and identification of rat fetal liver stem cells are simple and feasible.
【作者單位】： 南方醫(yī)科大學(xué)附屬解放軍第458醫(yī)院肝病防治中心;
【基金】：國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃(973計(jì)劃,2010CB945502)
【分類(lèi)號(hào)】：R329-33

【參考文獻(xiàn)】

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本文編號(hào)：1493780