Mechanical Stimulation Increases the Proliferation and Differentiation Potential of Human Adipose-Derived Stromal Cells

Jungwirth Susanne, Bohner Lauren, Spindler Kathrin, Hanisch Marcel, Kleinheinz Johannes, Sielker Sonja
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引用次数: 2

Abstract

Background: Dynamic conditions stimulate the bone remodeling process by improving the nutrients transport and increasing the expression of osteogenic cells. The purpose of this study was to evaluate the effect of mechanical stimulation on the osteogenic differentiation of human adipose-derived stromal cells. Methods: Cells were cultured under static and dynamic conditions in collagen scaffolds for 14 days. The mechanical stimulation was performed using a biaxial rotating bioreactor (5 × rpm and perfusion flow rate of 10 × rpm). Cell viability was analyzed with a living cell count and a MTT assay. Changes in expression of specific stem cell marker, osteogenic marker and endothelial markers were analyzed on gene (RT-qPCR) and protein (IHC) level. Data were statistically analyzed by one-way ANOVA (p = 0.05). Results: Cell viability was higher under dynamic condition and cells migrated deeper in the collagen matrix. Expression of stem cell marker (ANPEP/CD13, CD44, THY1/CD90) was significant higher under dynamic condition. This was also observed for osteogenic markers (collagen 1, osteopontin, osteonectin). Conclusion: The mechanical stimulation increased significantly cell viability and differentiation potential of human adipose-derived stromal cells.
机械刺激增加人脂肪来源基质细胞的增殖和分化潜能
背景:动态条件通过改善营养物质运输和增加成骨细胞的表达来刺激骨重塑过程。本研究旨在探讨机械刺激对人脂肪源性基质细胞成骨分化的影响。方法:细胞在胶原支架中静、动态培养14 d。机械刺激采用双轴旋转生物反应器(5 × rpm,灌注流速为10 × rpm)。用活细胞计数和MTT法分析细胞活力。从基因(RT-qPCR)和蛋白(IHC)水平分析特异性干细胞标志物、成骨标志物和内皮标志物的表达变化。资料采用单因素方差分析(p = 0.05)。结果:动态条件下细胞活力高,细胞在胶原基质中迁移深度大。动态条件下,干细胞标志物(ANPEP/CD13、CD44、THY1/CD90)的表达显著升高。在成骨标志物(胶原蛋白1、骨桥蛋白、骨粘连蛋白)中也观察到这一点。结论:机械刺激可显著提高人脂肪源性基质细胞的细胞活力和分化潜能。
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