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首页> 外文期刊>Journal of tissue engineering and regenerative medicine >Matrix stiffness and serum concentration effects matrix remodelling and ECM regulatory genes of human bone marrow stem cells
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Matrix stiffness and serum concentration effects matrix remodelling and ECM regulatory genes of human bone marrow stem cells

机译：基质刚度和血清浓度对人骨髓干细胞基质重构和ECM调控基因的影响

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The effects of mechanical stimulation of cell-seeded collagen constructs on cell orientation, intracellular signalling and molecular responses have been widely reported. In this study we investigated in vitro the contractile responses of human bone marrow stem cells (HBMSCs) to increasing collagen gel substrate stiffness and their effect on extracellular matrix (ECM) regulatory genes. Human dermal fibroblasts (HDFs) were used as controls. Cells were cultured in 10% and 20% FCS and embedded in collagen constructs at a density of 1 million cells/ml collagen. Matrix stiffness was achieved by subjecting the constructs to three different strain regimes (0%, 5% and 10%), using a computer-driven tensional culture force monitor (t-CFM) capable of uniaxial loading. The contraction forces generated by the cells were quantified over 24 h. Molecular outputs were quantified using RT-PCR. HBMSCs significantly increased force generation to increasing serum concentration (i.e 10% to 20%). 10% FCS concentration significantly reduced contraction as pre-strain stiffness was increased in HBMSCs and HDFs (0% > 5% > 10%). However, at 20% FCS HBMSCs generated similar peak force contraction at 24 h to 5% and 10% pre-strain (0% = 5% = 10%). The ECM regulatory gene for MMP2 showed upregulation at 5% pre-strain, but a 50% downregulation when pre-strain was increased to 10%. MMP9 was upregulated at 5% pre-strain and further upregulated at 10% pre-strain. In designing tissue-engineering solutions, predictable responses of cells, embedded within bio-artificial matrices, to external mechanical forces are critical. To take into account the increasing stiffness of the matrix as increasing ECM is deposited, it would be necessary to take mechanical stimulation into account to determine predictable cellular responses.
机译：机械接种细胞接种的胶原蛋白构造物对细胞取向，细胞内信号传导和分子反应的影响已被广泛报道。在这项研究中，我们在体外研究了人骨髓干细胞（HBMSC）对增加胶原蛋白凝胶底物刚度的收缩反应及其对细胞外基质（ECM）调节基因的影响。人皮肤成纤维细胞（HDF）用作对照。将细胞在10％和20％FCS中培养，并以100万细胞/ ml胶原的密度包埋在胶原构建物中。使用能够单轴加载的计算机驱动的张力培养力监控器（t-CFM），通过使构造体经受三种不同的应变方式（0％，5％和10％），可以实现基质刚度。在24小时内定量细胞产生的收缩力。使用RT-PCR定量分子输出。 HBMSC显着增加了产生力，从而增加了血清浓度（即10％至20％）。由于HBMSC和HDF的应变前刚度增加（10％> 5％> 10％），因此10％FCS浓度可显着减少收缩。然而，在FCS为20％时，HBMSC在24 h产生相似的峰值力收缩，分别为5％和10％的预应变（0％= 5％= 10％）。 MMP2的ECM调节基因在5％的预应变下显示上调，但在将预应变增加到10％时，下调了50％。 MMP9在5％的预应变上调，并在10％的预应变下进一步上调。在设计组织工程解决方案时，嵌入生物人工基质中的细胞对外部机械力的可预测响应至关重要。为了考虑到随着增加的ECM沉积而增加的基质刚度，有必要考虑机械刺激以确定可预测的细胞反应。

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