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猪肺脱细胞化以获得用于肺组织工程的三维支架。

Decellularization of Pig Lung to Yield Three-Dimensional Scaffold for Lung Tissue Engineering.

机构信息

Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.

International Clinical Research Center, St. Anne's University Hospital, Brno, Czech Republic.

出版信息

Methods Mol Biol. 2024;2764:21-33. doi: 10.1007/978-1-0716-3674-9_3.

Abstract

Lately, the need for three-dimensional (3D) cell culture has been recognized in order to closely mimic the organization of native tissues. Thus, 3D scaffolds started to be employed to facilitate the 3D cell organization and enable the artificial tissue formation for the emerging tissue engineering applications. 3D scaffolds can be prepared by various techniques, each with certain advantages and disadvantages. Decellularization is an easy method based on removal of cells from native tissue sample, yielding extracellular matrix (ECM) scaffold with preserved architecture and bioactivity. This chapter provides a detailed protocol for decellularization of pig lung and also some basic assays for evaluation of its effectivity, such as determination of DNA content and histological verification of the selected ECM components. Such decellularized scaffold can subsequently be used for various tissue engineering applications, for example, for recellularization with cells of interest, for natural ECM hydrogel preparation, or as a bioink for 3D bioprinting.

摘要

最近,人们认识到需要三维(3D)细胞培养,以便更紧密地模拟天然组织的结构。因此,3D 支架开始被用于促进 3D 细胞组织,并为新兴的组织工程应用实现人工组织形成。3D 支架可以通过各种技术制备,每种技术都有其优点和缺点。脱细胞化是一种基于从天然组织样本中去除细胞的简单方法,可产生具有保留的结构和生物活性的细胞外基质(ECM)支架。本章提供了一种详细的猪肺脱细胞化方案,以及一些用于评估其效果的基本检测方法,例如 DNA 含量的测定和选定 ECM 成分的组织学验证。这样的脱细胞支架随后可以用于各种组织工程应用,例如,用感兴趣的细胞再细胞化,用于天然 ECM 水凝胶的制备,或用作 3D 生物打印的生物墨水。

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