【摘要】：目的探究大鼠脂肪干細(xì)胞(r ADSCs)誘導(dǎo)成神經(jīng)細(xì)胞的可行性。方法第一步,通過(guò)酶消化法獲取r ADSCs并進(jìn)行培養(yǎng),通過(guò)CCK-8法檢測(cè)細(xì)胞增殖情況,并應(yīng)用免疫組化進(jìn)行鑒定,確定獲取的細(xì)胞為r ADSCs;第二步以原代培養(yǎng)的r ADSCs為研究對(duì)象,傳代至第2代后,采用神經(jīng)誘導(dǎo)培養(yǎng)基,誘導(dǎo)r ADSCs成神經(jīng)細(xì)胞,誘導(dǎo)所得細(xì)胞通過(guò)形態(tài)學(xué)及免疫熒光進(jìn)行鑒定。結(jié)果第一步中細(xì)胞培養(yǎng)增殖良好,增殖曲線(xiàn)顯示細(xì)胞生長(zhǎng)活躍,增殖能力強(qiáng);免疫組化檢測(cè)相關(guān)抗原CD90、CD105為陽(yáng)性,與造血干細(xì)胞相關(guān)抗原CD45為陰性,說(shuō)明所得細(xì)胞為r ADSCs。第二步中,誘導(dǎo)培養(yǎng)的細(xì)胞神經(jīng)球免疫熒光染色Nestin陽(yáng)性,細(xì)胞免疫熒光染色MAP-2陽(yáng)性,結(jié)合形態(tài)學(xué)可確定ADSCs已成功誘導(dǎo)分化成神經(jīng)細(xì)胞。結(jié)論 r ADSCs生長(zhǎng)活躍,表現(xiàn)出穩(wěn)定的增長(zhǎng)及增殖能力,并可定向誘導(dǎo)分化為神經(jīng)細(xì)胞。
[Abstract]:Objective to explore the feasibility of (r ADSCs) induction of rat adipose stem cells into neural cells. Methods in the first step, the r ADSCs was obtained by enzyme digestion and cultured. The proliferation of the cells was detected by CCK-8 method and identified by immunohistochemistry. The obtained cells were identified as r ADSCs;. In the second step, the primary cultured r ADSCs was used as the research object. After the passage to the second generation, the neurons of r ADSCs were induced by nerve induction medium. The induced cells were identified by morphology and immunofluorescence. Results in the first step, the cells were cultured and proliferated well, and the proliferation curve showed that the cells were active in growth and strong in proliferative ability, and that the relative antigen CD90,CD105 was positive and the CD45 associated with hematopoietic stem cells was negative, indicating that the cells obtained were r ADSCs.. In the second step, the cultured cells were stained with Nestin and MAP-2 with immunocytofluorescence staining. It was confirmed by morphology that ADSCs had been successfully induced to differentiate into nerve cells. Conclusion r ADSCs is active in growth, stable in growth and proliferation, and can be induced to differentiate into nerve cells.
【作者單位】： 成都軍區(qū)總醫(yī)院附屬口腔醫(yī)院;南充市中心醫(yī)院口腔科;
【基金】：四川省衛(wèi)生廳課題資助(120568)
【分類(lèi)號(hào)】：R329.2

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