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脱细胞去脂人胰腺衍生的细胞外基质支架和水凝胶。

Extracellular matrix scaffold and hydrogel derived from decellularized and delipidized human pancreas.

机构信息

Division of Transplantation, Department of Surgery, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, 53705, USA.

School of Pharmacy, University of Wisconsin, Madison, Wisconsin, 53705, USA.

出版信息

Sci Rep. 2018 Jul 11;8(1):10452. doi: 10.1038/s41598-018-28857-1.

Abstract

Extracellular matrix (ECM) plays an important developmental role by regulating cell behaviour through structural and biochemical stimulation. Tissue-specific ECM, attained through decellularization, has been proposed in several strategies for tissue and organ replacement. Decellularization of animal pancreata has been reported, but the same methods applied to human pancreas are less effective due to higher lipid content. Moreover, ECM-derived hydrogels can be obtained from many decellularized tissues, but methods have not been reported to obtain human pancreas-derived hydrogel. Using novel decellularization methods with human pancreas we produced an acellular, 3D biological scaffold (hP-ECM) and hydrogel (hP-HG) amenable to tissue culture, transplantation and proteomic applications. The inclusion of a homogenization step in the decellularization protocol significantly improved lipid removal and gelation capability of the resulting ECM, which was capable of gelation at 37 °C in vitro and in vivo, and is cytocompatible with a variety of cell types and islet-like tissues in vitro. Overall, this study demonstrates the characterisation of a novel protocol for the decellularization and delipidization of human pancreatic tissue for the production of acellular ECM and ECM hydrogel suitable for cell culture and transplantation applications. We also report a list of 120 proteins present within the human pancreatic matrisome.

摘要

细胞外基质 (ECM) 通过结构和生化刺激调节细胞行为,在发育过程中发挥着重要作用。通过脱细胞化获得的组织特异性 ECM 已在几种组织和器官替代策略中得到提出。已经有报道称对动物胰腺进行脱细胞处理,但由于脂质含量较高,同样的方法在人胰腺中效果较差。此外,许多脱细胞组织都可以获得 ECM 衍生的水凝胶,但尚未有报道称可以获得人胰腺衍生的水凝胶。我们使用新的人胰腺脱细胞方法,制备了一种无细胞的 3D 生物支架 (hP-ECM) 和水凝胶 (hP-HG),适用于组织培养、移植和蛋白质组学应用。在脱细胞方案中包含均质化步骤,可显著提高 ECM 的脂质去除和凝胶化能力,从而使其能够在 37°C 下进行体外和体内凝胶化,并且与多种细胞类型和体外胰岛样组织具有细胞相容性。总体而言,这项研究展示了一种新型的人胰腺组织脱细胞和去脂化方案,用于制备适用于细胞培养和移植应用的无细胞 ECM 和 ECM 水凝胶。我们还报告了人类胰腺基质组中存在的 120 种蛋白质列表。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1173/6041318/36063eea8395/41598_2018_28857_Fig1_HTML.jpg

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