動態(tài)壓應(yīng)力與共培養(yǎng)微環(huán)境對脂肪間充質(zhì)干細(xì)胞增殖及向髓核樣細(xì)胞分化的影響
發(fā)布時間：2018-08-03 07:11
【摘要】：【目的】探討動態(tài)壓應(yīng)力與共培養(yǎng)微環(huán)境條件對大鼠脂肪間充質(zhì)干細(xì)胞（ADSCs）增殖及向髓核樣細(xì)胞分化的影響。 【方法】取原代SD大鼠ADSCs體外培養(yǎng)并擴(kuò)增，流式細(xì)胞儀鑒定細(xì)胞表面標(biāo)志物后，以第三代ADSCs經(jīng)可控細(xì)胞動態(tài)壓應(yīng)力加載裝置(90mmHg，12h/d)行刺激，并設(shè)置陰性組，采用CFSE染色法測細(xì)胞增殖活性，分別于染色后24h,48h,72h，96h以倒置熒光顯微鏡觀察并以流式細(xì)胞儀檢測熒光衰減度。同時將包裹有SD大鼠髓核細(xì)胞（NPCs）的藻酸鹽微球與第三代ADSCs混合共培養(yǎng)，按細(xì)胞共培養(yǎng)和壓應(yīng)力刺激與否分為單純ADSCs培養(yǎng)組、ADSCs加壓組、單純NPCs培養(yǎng)組、NPCs加壓組、共培養(yǎng)組和共培養(yǎng)聯(lián)合壓力組，于干預(yù)至第7d時采用RealtimePCR和WesternBlot檢測各組貼壁細(xì)胞SOX-9，Ⅱ型膠原及aggrecan的表達(dá)差異。 【結(jié)果】實(shí)驗(yàn)所用ADSCs經(jīng)流式細(xì)胞儀鑒定顯示CD29、CD44陽性率分別為99.72%、99.55%，，CD34、CD45陽性率分別為1.40%、1.80%；CFSE結(jié)果顯示ADSCs壓力組細(xì)胞的增殖活性較單純ADSCs培養(yǎng)組顯著提高；軟骨特異性指標(biāo)SOX-9，Ⅱ型膠原及aggrecan的基因表達(dá)水平在共培養(yǎng)聯(lián)合壓力組中增加最為顯著，其中Ⅱ型膠原基因表達(dá)水平變化與其蛋白表達(dá)量變化相一致。 【結(jié)論】動態(tài)壓應(yīng)力能促進(jìn)ADSCs的增殖，并且在聯(lián)合NPCs共培養(yǎng)微環(huán)境下可誘導(dǎo)ADSCs向髓核樣細(xì)胞分化。
[Abstract]:[objective] to investigate the effects of dynamic compressive stress and co-culture microenvironment on the proliferation and differentiation of adipose mesenchymal stem cells (ADSCs) into nucleus pulposus like cells. [methods] Primary SD rat ADSCs was cultured and amplified in vitro. After identification of cell surface markers by flow cytometry, the third generation of ADSCs was stimulated by controllable cell dynamic compressive stress loading device (90 mm Hg + 12 h / d), and negative group was set up. Cell proliferation activity was measured by CFSE staining. The fluorescence attenuation was detected by flow cytometry and inverted fluorescence microscope was used to observe the fluorescence attenuation at 24 hours after dyeing for 48 h and 72 h for 96 h. At the same time, the alginate microspheres coated with (NPCs) of SD rat nucleus pulposus were co-cultured with ADSCs of the third generation. According to the cell co-culture and compressive stress stimulation or not, they were divided into ADSCs compression group and NPCs culture group. Co-culture group and co-culture combined pressure group, The expression of SOX-9, type 鈪
本文編號：2160992
[Abstract]:[objective] to investigate the effects of dynamic compressive stress and co-culture microenvironment on the proliferation and differentiation of adipose mesenchymal stem cells (ADSCs) into nucleus pulposus like cells. [methods] Primary SD rat ADSCs was cultured and amplified in vitro. After identification of cell surface markers by flow cytometry, the third generation of ADSCs was stimulated by controllable cell dynamic compressive stress loading device (90 mm Hg + 12 h / d), and negative group was set up. Cell proliferation activity was measured by CFSE staining. The fluorescence attenuation was detected by flow cytometry and inverted fluorescence microscope was used to observe the fluorescence attenuation at 24 hours after dyeing for 48 h and 72 h for 96 h. At the same time, the alginate microspheres coated with (NPCs) of SD rat nucleus pulposus were co-cultured with ADSCs of the third generation. According to the cell co-culture and compressive stress stimulation or not, they were divided into ADSCs compression group and NPCs culture group. Co-culture group and co-culture combined pressure group, The expression of SOX-9, type 鈪
本文編號：2160992
本文鏈接：http://sikaile.net/xiyixuelunwen/2160992.html
最近更新
教材專著
