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Bioreactor-induced mesenchymal progenitor cell differentiation and elastic fiber assembly in engineered vascular tissues

机译:生物反应器诱导的间充质祖细胞分化与工程血管组织中的弹性纤维组件

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In vitro maturation of engineered vascular tissues (EVT) requires the appropriate incorporation of smooth muscle cells (SMC) and extracellular matrix (ECM) components similar to native arteries. To this end, the aim of the current study was to fabricate 4 mm inner diameter vascular tissues using mesenchymal progenitor cells seeded into tubular scaffolds. A dual-pump bioreactor operating either in perfusion or pulsatile perfusion mode was used to generate physiological-like stimuli to promote progenitor cell differentiation, extracellular elastin production, and tissue maturation. Our data demonstrated that pulsatile forces and perfusion of 3D tubular constructs from both the lumenal and ablumenal sides with culture media significantly improved tissue assembly, effectively inducing mesenchymal progenitor cell differentiation to SMCs with contemporaneous elastin production. With bioreactor cultivation, progenitor cells differentiated toward smooth muscle lineage characterized by the expression of smooth muscle (SM)-specific markers smooth muscle alpha actin (SM-alpha-actin) and smooth muscle myosin heavy chain (SM-MHC). More importantly, pulsatile perfusion bioreactor cultivation enhanced the synthesis of tropoelastin and its extracellular cross-linking into elastic fiber compared with static culture controls. Taken together, the current study demonstrated progenitor cell differentiation and vascular tissue assembly, and provides insights into elastin synthesis and assembly to fibers.
机译:工程化血管组织(EVT)的体外成熟需要适当掺入与天然动脉相似的平滑肌细胞(SMC)和细胞外基质(ECM)组分。为此,目前研究的目的是使用播种到管状支架中的间充质祖细胞来制造4mm内径的血管组织。在灌注或脉冲灌注模式下操作的双泵生物反应器用于产生类似的生理刺激以促进祖细胞分化,细胞外弹性蛋白产生和组织成熟。我们的数据表明,来自培养培养介质的脉络和3D管状构建体的脉动力和3D管状构建体的灌注显着改善了组织组件,有效地诱导了具有同期弹性蛋白产生的间充质祖细胞对SMC的分化。伴有生物反应器培养,祖细胞分化为平滑肌谱系,其特征在于平滑肌(SM)的表达式 - 特异性标志物平滑肌α肌动蛋白(SM-Alpha-Actin)和平滑肌肌霉素重链(SM-MHC)。更重要的是,与静态培养对照相比,脉动灌注生物反应器培养增强了Tropoelastin的合成及其细胞外交联进入弹性纤维。在一起,目前的研究表明祖细胞分化和血管组织组件,并向Elastin合成和组装提供给纤维的见解。

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