【摘要】：目的探討大鼠骨髓間充質(zhì)干細(xì)胞(BMSC)對(duì)大鼠RSC96施萬(wàn)細(xì)胞增殖及遷移的影響。方法全骨髓細(xì)胞貼壁法分離并提取SD大鼠BMSC,倒置顯微鏡觀察第3代BMSC形態(tài);流式細(xì)胞術(shù)檢測(cè)細(xì)胞表面CD19、CD34、CD73、CD105;第3代BMSC經(jīng)成脂成骨誘導(dǎo)液分別誘導(dǎo)3周和2周后,分別應(yīng)用油紅O染色和堿性磷酸酶檢測(cè)BMSC的多向分化能力。應(yīng)用24 mm共培養(yǎng)板,將第3代BMSC與RSC96細(xì)胞共培養(yǎng)24 h,采用MTT法和平板克隆形成實(shí)驗(yàn)檢測(cè)RSC96細(xì)胞增殖及克隆形成能力;劃痕實(shí)驗(yàn)及TranswellTM小室實(shí)驗(yàn)檢測(cè)RSC96細(xì)胞的遷移能力;Western blot法檢測(cè)RSC96細(xì)胞Bax和Bcl-2蛋白表達(dá)水平。結(jié)果SD大鼠第3代BMSC顯微鏡下呈梭形、旋渦狀排列,形態(tài)大小相似;BMSC表面標(biāo)志CD73、CD105高表達(dá),造血干細(xì)胞表面標(biāo)志CD34和淋巴細(xì)胞表面標(biāo)志CD19低表達(dá);BMSC經(jīng)成脂誘導(dǎo)培養(yǎng)3周后,細(xì)胞有大量脂滴;經(jīng)成骨誘導(dǎo)培養(yǎng)2周后,堿性磷酸酶染色呈陽(yáng)性;BMSC與RSC96細(xì)胞共培養(yǎng)后,與對(duì)照組相比,RSC96細(xì)胞的增殖活性顯著降低,克隆形成能力顯著減弱、細(xì)胞遷移能力降低;共培養(yǎng)后RSC96細(xì)胞Bax蛋白水平升高,Bcl-2蛋白表達(dá)降低。結(jié)論 BMSC能促進(jìn)RSC96細(xì)胞凋亡,抑制其增殖、遷移。
[Abstract]:Objective to investigate the effect of rat bone marrow mesenchymal stem cell (BMSC) on proliferation and migration of rat RSC96 Schwann cells. Methods BMSC, of SD rats was isolated and extracted by whole bone marrow adherent method. The morphology of the third generation BMSC was observed by inverted microscope, and the third generation of BMSC on the cell surface was detected by flow cytometry after being induced by adipogenic osteoblasts for 3 weeks and 2 weeks, respectively. Oil red O staining and alkaline phosphatase were used to detect the multidirectional differentiation ability of BMSC. The third generation of BMSC and RSC96 cells were co-cultured for 24 h with 24 mm co-culture plate. The proliferation and clone forming ability of RSC96 cells were detected by MTT assay and plate clone formation assay. The migration ability of RSC96 cells was detected by scratch test and TranswellTM chamber assay. The expression of Bax and Bcl-2 in RSC96 cells was detected by; Western blot method. Results the third generation BMSC of SD rats showed spindle shape, swirl arrangement and similar size under microscope, high expression of CD73,CD105 on BMSC surface, low expression of CD34 on hematopoietic stem cell surface and CD19 on lymphocyte surface, BMSC was induced by adipogenic culture for 3 weeks. After 2 weeks of osteogenic induction, alkaline phosphatase staining was positive. After co-culture of BMSC and RSC96 cells, the proliferation activity of RSC96 cells was significantly lower than that of the control group, and the ability of clone formation was significantly decreased. The level of Bax protein increased and the expression of Bcl-2 protein decreased after co-culture. Conclusion BMSC can promote apoptosis, inhibit proliferation and migration of RSC96 cells.
【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) 常州市衛(wèi)生局重大科研立項(xiàng)(ZD201508)
【分類號(hào)】：R329.2
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本文編號(hào)：2272313