用于软骨再生的脱细胞细胞外基质支架

Decellularized Extracellular Matrix Scaffolds for Cartilage Regeneration.

作者信息

Thakkar Shraddha, Fernandes Hugo, Moroni Lorenzo

机构信息

Department of Biomedical Engineering, Eindhoven University of Technology, 513, 5600MB, Eindhoven, The Netherlands.

Tissue Regeneration Department, University of Maastricht - MERLN Institute for Technology Inspired Regenerative Medicine Complex, Tissue Regeneration Department Universiteitsingel 40, 6229 ER, Maastricht, The Netherlands.

出版信息

Methods Mol Biol. 2015;1340:133-51. doi: 10.1007/978-1-4939-2938-2_9.

DOI:10.1007/978-1-4939-2938-2_9

PMID:26445835

Abstract

Decellularized extracellular matrix (ECM) is gaining a lot of attention as a biomaterial for tissue engineering applications. This chapter describes the processing techniques for decellularization of cell-derived ECM and protocols for the fabrication of ECM-based scaffolds in the form of hydrogels or fibrous polymer meshes by electrospinning. It describes the protocols to analyze the morphology and presence of collagen in fabricated scaffolds using scanning electron microscope and Picrosirius Red staining respectively. Methods to evaluate the metabolic activity and proliferation of cells (resazurin-based assay and DNA assay, respectively) and gene expression are also presented. Furthermore, histological techniques to analyze the presence of sulfated glycosaminoglycans are also described.

摘要

去细胞化细胞外基质（ECM）作为一种用于组织工程应用的生物材料正受到广泛关注。本章介绍了细胞来源ECM的去细胞化处理技术，以及通过静电纺丝制备水凝胶或纤维聚合物网形式的基于ECM的支架的方案。它描述了分别使用扫描电子显微镜和天狼星红染色分析制备支架中胶原蛋白的形态和存在情况的方案。还介绍了评估细胞代谢活性和增殖的方法（分别基于刃天青的测定法和DNA测定法）以及基因表达。此外，还描述了分析硫酸化糖胺聚糖存在情况的组织学技术。