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Isolation and identification of extracellular matrix proteins from oil-based CASPERized mouse brains for matrisomal analysis

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Title
Isolation and identification of extracellular matrix proteins from oil-based CASPERized mouse brains for matrisomal analysis
Author(s)
Ha, Byung Geun; Jang, Yu-Jin; Lee, EunSoo; Byung-Gyu Kim; Kyungjae Myung; Sun, Woong; Jeong, Sung-Jin
Publication Date
2023-04
Journal
Heliyon, v.9, no.4
Publisher
Elsevier Ltd
Abstract
The extracellular matrix (ECM) components present within all tissues and organs help to maintain the cytoskeletal architecture and tissue morphology. Although the ECM plays a role in cellular events and signaling pathways, it has not been well studied due its insolubility and complexity. Brain tissue has a higher cell density and weaker mechanical strength than other tissues in the body. When removing cells using a general decellularization method to produce scaffolds and obtain ECM proteins, various problems must be considered because tissues are easily damaged. To retain the brain shape and ECM components, we performed decellularization in combination with polymerization. We immersed mouse brains in oil for polymerization and decellularization via O-CASPER (Oil-based Clinically and Experimentally Applicable Acellular Tissue Scaffold Production for Tissue Engineering and Regenerative Medicine) and then isolated ECM components using sequential matrisome preparation reagents (SMPRs), namely, RIPA, PNGase F, and concanavalin A. Adult mouse brains were preserved with our decellularization method. Western blot and LC-MS/MS analyses revealed that ECM components, including collagen and laminin, were isolated efficiently from decellularized mouse brains using SMPRs. Our method will be useful to obtain matrisomal data and perform functional studies using adult mouse brains and other tissues. © 2023
URI
https://pr.ibs.re.kr/handle/8788114/13640
DOI
10.1016/j.heliyon.2023.e14777
ISSN
2405-8440
Appears in Collections:
Center for Genomic Integrity(유전체 항상성 연구단) > 1. Journal Papers (저널논문)
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