{"title":"Decellularized Extracellular Matrix for Cell Biology.","authors":"Takashi Hoshiba","doi":"10.1002/cpz1.318","DOIUrl":null,"url":null,"abstract":"<p><p>The extracellular matrix (ECM) is an architecture that supports the cells in our bodies and regulates various cell functions. The ECM is composed of many proteins and carbohydrates, and these molecules activate various intracellular signaling pathways orchestrated to decide cell fates. Therefore, it is not enough to study the role of single ECM molecules to understand the roles of the ECM in the regulation of cell functions; it is necessary to understand how the ECM, as an assembly of various molecules, regulates cell functions as a whole. For this purpose, in vitro ECM models mimicking native ECM are required. Here, a decellularization technique is presented to reconstitute native ECM in vitro. In this article, methods for preparing decellularized ECM (dECM) are described for use in tumor and stem cell biology. Additionally, a method for confirmation of decellularization and a dECM modification method are described. These dECM types will be useful for comprehensive studies of ECM roles. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Preparation of in vitro extracellular matrix (ECM) models mimicking native ECM in different malignant tumor tissues Basic Protocol 2: Preparation of in vitro ECM models mimicking native ECM surrounding myoblasts differentiating into myotubes at each myogenic stage Support Protocol 1: Confirmation of myogenic stages by myogenic stages by myogenic gene expression analysis Basic Protocol 3: Confirmation of cell removal Basic Protocol 4: Reduction of chondroitin sulfate chains in cultured cell-derived decellularized ECM Support Protocol 2: Quantification of chondroitin sulfate chain amounts in the decellularized ECM.</p>","PeriodicalId":11174,"journal":{"name":"Current Protocols","volume":" ","pages":"e318"},"PeriodicalIF":0.0000,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"3","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cpz1.318","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3
Abstract
The extracellular matrix (ECM) is an architecture that supports the cells in our bodies and regulates various cell functions. The ECM is composed of many proteins and carbohydrates, and these molecules activate various intracellular signaling pathways orchestrated to decide cell fates. Therefore, it is not enough to study the role of single ECM molecules to understand the roles of the ECM in the regulation of cell functions; it is necessary to understand how the ECM, as an assembly of various molecules, regulates cell functions as a whole. For this purpose, in vitro ECM models mimicking native ECM are required. Here, a decellularization technique is presented to reconstitute native ECM in vitro. In this article, methods for preparing decellularized ECM (dECM) are described for use in tumor and stem cell biology. Additionally, a method for confirmation of decellularization and a dECM modification method are described. These dECM types will be useful for comprehensive studies of ECM roles. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Preparation of in vitro extracellular matrix (ECM) models mimicking native ECM in different malignant tumor tissues Basic Protocol 2: Preparation of in vitro ECM models mimicking native ECM surrounding myoblasts differentiating into myotubes at each myogenic stage Support Protocol 1: Confirmation of myogenic stages by myogenic stages by myogenic gene expression analysis Basic Protocol 3: Confirmation of cell removal Basic Protocol 4: Reduction of chondroitin sulfate chains in cultured cell-derived decellularized ECM Support Protocol 2: Quantification of chondroitin sulfate chain amounts in the decellularized ECM.
用于细胞生物学的脱细胞细胞外基质。
细胞外基质(ECM)是一种支持我们体内细胞并调节各种细胞功能的结构。ECM由许多蛋白质和碳水化合物组成,这些分子激活各种细胞内信号通路,以决定细胞命运。因此,仅研究单个ECM分子的作用不足以了解ECM在调节细胞功能中的作用;有必要了解ECM作为各种分子的集合,如何作为一个整体调节细胞功能。为此,需要模拟天然ECM的体外ECM模型。在这里,提出了一种脱细胞技术来体外重建天然ECM。本文介绍了用于肿瘤和干细胞生物学的脱细胞ECM (dECM)的制备方法。此外,还描述了一种确认脱细胞的方法和一种dECM修饰方法。这些dECM类型将有助于ECM角色的全面研究。©2021 Wiley期刊有限责任公司基本方案1:制备体外细胞外基质(ECM)模型,模拟不同恶性肿瘤组织中的天然ECM。基本方案2:制备体外ECM模型,模拟成肌细胞周围的天然ECM,在每个肌生成阶段分化为肌管。支持方案1:通过肌生成基因表达分析,通过肌生成阶段确认肌生成阶段。基本方案3:确认细胞移除。基本方案4:在培养细胞来源的脱细胞ECM中硫酸软骨素链的减少支持方案2:定量测定脱细胞ECM中硫酸软骨素链的数量。
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