【摘要】：目的:探討大鼠骨髓間充質(zhì)干細(xì)胞(BMSC)培養(yǎng)上清液對大鼠RSC96細(xì)胞生物學(xué)功能的影響。方法:體外培養(yǎng)、純化SD大鼠BMSC并進(jìn)行鑒定;收集第3代BMSC培養(yǎng)上清液(BMSC-CM);采用MTT法檢測BMSC-CM對RSC96細(xì)胞增殖的影響;平板克隆實(shí)驗(yàn)分析BMSC-CM對RSC96單個(gè)細(xì)胞增殖的影響;劃痕實(shí)驗(yàn)和TranswellTM小室實(shí)驗(yàn)分析BMSC-CM對RSC96細(xì)胞遷移的作用,Western blot法檢測BMSC-CM對RSC96細(xì)胞Bax和Bcl-2蛋白表達(dá)的改變。結(jié)果:SD大鼠第3代BMSC形態(tài)呈長梭形,旋渦樣生長;成骨染色為陽性結(jié)果;成脂誘導(dǎo)后有脂滴出現(xiàn);BMSC-CM作用后抑制RSC96細(xì)胞的增殖和遷移能力,且作用后的RSC96細(xì)胞Bax蛋白水平升高,Bcl-2蛋白表達(dá)降低。結(jié)論:BMSC-CM促進(jìn)RSC96細(xì)胞凋亡,抑制其增殖和遷移。
[Abstract]:Objective: To study the effect of culture supernatant of rat bone marrow mesenchymal stem cells (BMSC) on the biological function of rat RSC96 cells. Methods: In vitro culture, the BMSC of SD rats was purified and identified; the third generation BMSC was collected to culture supernatant (BMSC-CM); the effect of BMSC-CM on the proliferation of RSC96 cells was detected by MTT method; the effect of BMSC-CM on the proliferation of single cell of RSC96 was analyzed by plate cloning. The effect of BMSC-CM on the cell migration of the RSC96 cells was analyzed by the scratch test and the Transwell (TM) cell experiment. The expression of Bax and Bcl-2 in the cells of the RSC96 cells was detected by Western blot. Results: The morphology of the third generation BMSC of SD rats was in the form of long-shuttle and vortex-like growth, the positive results were found in the bone formation, and after the formation of lipid, the proliferation and migration of the RSC96 cells were inhibited, and the level of Bax protein in the RSC96 cells after the action was increased, and the expression of Bcl-2 protein was decreased. Conclusion: BMSC-CM promotes the apoptosis of RSC96 cells and inhibits its proliferation and migration.
【作者單位】： 江蘇大學(xué)附屬金壇醫(yī)院;江蘇大學(xué)附屬醫(yī)院;
【基金】：江蘇省自然科學(xué)基金(BK20141295) 江蘇大學(xué)醫(yī)學(xué)臨床科技發(fā)展基金項(xiàng)目(JLY20160081) 江蘇省“六個(gè)一”工程項(xiàng)目(LGY2016055)
【分類號(hào)】：R392
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本文編號(hào)：2491877