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Comparison of Proliferation and Osteogenic Differentiation Potential of Rat Mandibular and Femoral Bone Marrow Mesenchymal Stem Cells In Vitro

机译:大鼠颌骨骨髓间充质干细胞的增殖和骨质骨髓间充质干细胞的增殖和骨质发生分化潜力的比较

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This study was conducted to compare the in vitro proliferation and osteogenic differentiation potential of mesenchymal stem cells (MSCs) derived from mandibular (M-MSCs) or femur (F-MSCs) tissues of rats. M-MSC and F-MSC cultures were isolated and established from the same rat. Cultures were observed for morphological changes by microscope and growth characteristics by CCK-8 and cloning assays. Cell adhesion ability on a culture plate and titanium sheet was detected by staining with toluidine blue and Hoechst 33258, respectively. The levels of Ca, P, and ALP (serially) during osteogenic differentiation were evaluated. Cultures were analyzed for mineralization potential with alizarin red and ALP staining methods and for differentiation markers with RT-PCR (ALP, Runx2, and OCN). M-MSCs and F-MSCs were successfully isolated from the same rat with uncontaminated culture, which showed significant differences in morphology. The proliferation rate of M-MSCs was higher than F-MSCs in primary culture, but significantly lower after passage. More colonies are formed from F-MSCs than from M-MSCs. M-MSCs showed a significantly higher mineralization and osteogenic differentiation potential, which might be of significance for use in bone/dental tissue engineering. In vitro, cell passage will decrease the proliferation ability of M-MSCs. The higher mineralization and osteogenic differentiation potential of M-MSCs could make them an approachable stem cell source for further application in stem cell-based clinical therapies.
机译:进行该研究以比较衍生自大鼠颌骨(MSCs)或股骨(F-MSCs)组织的间充质干细胞(MSCs)的体外增殖和成骨分化电位。分离M-MSC和F-MSC培养物,并从同一大鼠建立。通过CCK-8和克隆测定观察到通过显微镜和生长特性的形态变化而观察到培养物。通过用甲苯胺蓝和Hoechst 33258染色,检测培养板和钛板上的细胞粘附能力。评估了骨质发生分化期间Ca,P和Alp(连续)的水平。用茜素红和ALP染色方法和具有RT-PCR的分化标志物(ALP,RUNX2和OCN)分析培养物进行矿化电位。 M-MSCs和F-MSCs与不同的大鼠用不同污染的培养成功分离出来,这表明了形态学的显着差异。 M-MSCs的增殖率高于初级培养物中的F-MSC,但通过后显着降低。更多来自F-MSCs的菌落而不是M-MSCs。 M-MSCs显示出明显更高的矿化和成骨分化电位,这可能具有在骨/牙科组织工程中使用的重要性。体外,细胞通过将降低MSCs的增殖能力。 M-MSCs的更高的矿化和成骨分化电位可以使它们成为可接近的干细胞源,用于进一步应用于干细胞的临床疗法。

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