While extracellular matrix (ECM)-derived coatings have the potential to direct the response of cell populations in culture, there is a need to investigate the effects of ECM sourcing and processing on substrate bioactivity. To develop improved cell culture models for studying adipogenesis, the current study examines the proliferation and adipogenic differentiation of human adipose-derived stem/stromal cells (ASCs) on a range of ECM-derived coatings. Human decellularized adipose tissue (DAT) and commercially available bovine tendon collagen (COL) are digested with α-amylase or pepsin to prepare the coatings. Physical characterization demonstrates that α-amylase digestion generates softer, thicker, and more stable coatings, with a fibrous tissue-like ultrastructure that is lost in the pepsin-digested thin films. ASCs cultured on the α-amylase-digested ECM have a more spindle-shaped morphology, and proliferation is significantly enhanced on the α-amylase-digested DAT coatings. Further, the α-amylase-digested DAT provides a more pro-adipogenic microenvironment, based on higher levels of adipogenic gene expression, glycerol-3-phosphate dehydrogenase (GPDH) enzyme activity, and perilipin staining. Overall, this study supports α-amylase digestion as a new approach for generating bioactive ECM-derived coatings, and demonstrates tissue-specific bioactivity using adipose-derived ECM to enhance ASC proliferation and adipogenic differentiation.

中文翻译：

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在α-淀粉酶消化的脱细胞脂肪组织衍生的组织特异性涂层上进行培养可增强人脂肪衍生的基质细胞的增殖和成脂分化。

尽管细胞外基质（ECM）衍生的涂层具有指导培养中细胞群体反应的潜力，但仍需要研究ECM来源和加工对基质生物活性的影响。为了开发用于研究脂肪形成的改进的细胞培养模型，当前的研究检查了人类脂肪来源的干/基质细胞（ASC）在一系列ECM来源的涂层上的增殖和脂肪形成分化。用α-淀粉酶或胃蛋白酶消化人脱细胞的脂肪组织（DAT）和市售的牛腱胶原（COL）以制备涂层。物理特征表明，α-淀粉酶消化可产生更柔软，更厚和更稳定的涂层，并具有在胃蛋白酶消化的薄膜中消失的纤维组织状超微结构。在α-淀粉酶消化的ECM上培养的ASC具有更多的纺锤形形态，并且在α-淀粉酶消化的DAT涂层上增殖显着增强。此外，基于更高水平的成脂基因表达，3-磷酸甘油三磷酸脱氢酶（GPDH）酶活性和脂蛋白染色，α-淀粉酶消化的DAT提供了更有利于成脂的微环境。总的来说，这项研究支持α-淀粉酶消化作为产生具有生物活性的ECM的涂层的新方法，并证明了使用脂肪来源的ECM增强ASC增殖和成脂分化的组织特异性生物活性。基于更高水平的成脂基因表达，3-磷酸甘油脱氢酶（GPDH）酶活性和脂蛋白染色。总的来说，这项研究支持α-淀粉酶消化作为产生具有生物活性的ECM的涂层的新方法，并证明了使用脂肪来源的ECM增强ASC增殖和成脂分化的组织特异性生物活性。基于更高水平的成脂基因表达，3-磷酸甘油脱氢酶（GPDH）酶活性和脂蛋白染色。总的来说，这项研究支持α-淀粉酶消化作为产生具有生物活性的ECM的涂层的新方法，并证明了使用脂肪来源的ECM增强ASC增殖和成脂分化的组织特异性生物活性。