The effect of mechanical stress on the proliferation, adipogenic differentiation and gene expression of human adipose‐derived stem cells
The effect of mechanical stress on the proliferation, adipogenic differentiation and gene expression of human adipose‐derived stem cells
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DOI：
10.1002/term.2411
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发表时间：
2018-01
影响因子：
3.3
通讯作者：
N. Paul;B. Denecke;Bong-Sung Kim;A. Dreser;J. Bernhagen;N. Pallua
中科院分区：
文献类型：
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作者：
N. Paul;B. Denecke;Bong-Sung Kim;A. Dreser;J. Bernhagen;N. Pallua
为了更好地将外部体积扩张应用于软组织再生的临床应用，有必要全面了解其潜在机制。由于人类脂肪干细胞（hASC）在软组织扩张中起着至关重要的作用，我们研究了周期性拉伸对这些细胞的基因表达，增殖率和成脂分化的影响。循环拉伸后，提取RNA并进行DNA微阵列分析和逆转录-定量聚合酶链反应（RT-qPCR）。此外，通过RT-qPCR分析FABP 4 mRNA的表达，以测试机械拉伸是否影响hASC的成脂分化。使用alamarBlue测定和Ki-67染色评估增殖率。流式细胞术和Western blot分析细胞周期。我们发现，周期性牵张显著诱导CYP 1B 1 mRNA的表达。此外，hASCs的成脂分化受损，增殖也受损。这部分是由于细胞外信号调节激酶（ERK）1/2和组蛋白H3磷酸化的减少，表明细胞周期的G2/M期生长停滞。富集分析表明，拉伸调节基因在参与细胞外基质组织，血管重塑和细胞应激反应的途径和生物过程中过度表达。总之，机械应力损害了增殖和成脂分化，但导致了hASC的组织重塑表型。这些数据表明，细胞外基质重塑和新血管生成可能发挥更重要的作用，在外部体积扩张比增殖和脂肪细胞的。版权所有© 2017约翰威利父子有限公司
To allow for a better implementation of external volume expansion to clinical applications for soft tissue regeneration, it is necessary to comprehensively understand the underlying mechanisms. As human adipose‐derived stem cells (hASCs) play a crucial role in soft tissue enlargement, we investigated the impact of cyclic stretch on gene expression, proliferation rate and adipogenic differentiation of these cells. After cyclic stretching, RNA was extracted and subjected to DNA microarray analysis and reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR). Also, the expression of FABP4 mRNA was analysed by RT‐qPCR to test whether mechanical stretch affected adipogenic differentiation of hASCs. The proliferation rate was assessed using the alamarBlue assay and Ki‐67 staining. A cell cycle analysis was performed with flow cytometry and Western blot. We found that cyclic stretch significantly induced the expression of CYP1B1 mRNA. Furthermore, the adipogenic differentiation of hASCs was impaired, as was the proliferation. This was partly due to a decrease in extracellular signal‐regulated kinase (ERK) 1/2 and histone H3 phosphorylation, suggesting a growth arrest in the G2/M phase of the cell cycle. Enrichment analyses demonstrated that stretch‐regulated genes were over‐represented in pathways and biological processes involved in extracellular matrix organization, vascular remodelling and responses to cell stress. Taken together, mechanical stress impaired both proliferation and adipogenic differentiation, but led to a tissue‐remodelling phenotype of hASCs. These data suggest that extracellular matrix remodelling and neoangiogenesis may play a more important role in external volume expansion than proliferation and adipogenesis of hASCs. Copyright © 2017 John Wiley & Sons, Ltd.