Universität zu Köln

Mayorca-Guiliani, Alejandro E., Willacy, Oliver ORCID: 0000-0002-1541-7795, Madsen, Chris D., Rafaeva, Maria, Heumueller, Stefanie Elisabeth, Bock, Felix, Sengle, Gerhard, Koch, Manuel, Imhof, Thomas, Zaucke, Frank ORCID: 0000-0002-7680-9354, Wagener, Raimund, Sasaki, Takako, Erler, Janine T. and Reuten, Raphael (2019). Decellularization and antibody staining of mouse tissues to map native extracellular matrix structures in 3D. Nat. Protoc., 14 (12). S. 3395 - 3426. LONDON: NATURE PUBLISHING GROUP. ISSN 1750-2799

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Abstract

The extracellular matrix (ECM) is a major regulator of homeostasis and disease, yet the 3D structure of the ECM remains poorly understood because of limitations in ECM visualization. We recently developed an ECM-specialized method termed in situ decellularization of tissues (ISDoT) to isolate native 3D ECM scaffolds from whole organs in which ECM structure and composition are preserved. Here, we present detailed surgical instructions to facilitate decellularization of 33 different mouse tissues and details of validated antibodies that enable the visualization of 35 mouse ECM proteins. Through mapping of these ECM proteins, the structure of the ECM can be determined and tissue structures visualized in detail. In this study, perfusion decellularization is presented for bones, skeletal muscle, tongue, salivary glands, stomach, duodenum, jejunum/ileum, large intestines, mesentery, liver, gallbladder, pancreas, trachea, bronchi, lungs, kidneys, urinary bladder, ovaries, uterine horn, cervix, adrenal gland, heart, arteries, veins, capillaries, lymph nodes, spleen, peripheral nerves, eye, outer ear, mammary glands, skin, and subcutaneous tissue. Decellularization, immunostaining, and imaging take 4-5 d.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Mayorca-Guiliani, Alejandro E. UNSPECIFIED UNSPECIFIED UNSPECIFIED Willacy, Oliver UNSPECIFIED orcid.org/0000-0002-1541-7795 UNSPECIFIED Madsen, Chris D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Rafaeva, Maria UNSPECIFIED UNSPECIFIED UNSPECIFIED Heumueller, Stefanie Elisabeth UNSPECIFIED UNSPECIFIED UNSPECIFIED Bock, Felix UNSPECIFIED UNSPECIFIED UNSPECIFIED Sengle, Gerhard UNSPECIFIED UNSPECIFIED UNSPECIFIED Koch, Manuel UNSPECIFIED UNSPECIFIED UNSPECIFIED Imhof, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Zaucke, Frank UNSPECIFIED orcid.org/0000-0002-7680-9354 UNSPECIFIED Wagener, Raimund UNSPECIFIED UNSPECIFIED UNSPECIFIED Sasaki, Takako UNSPECIFIED UNSPECIFIED UNSPECIFIED Erler, Janine T. UNSPECIFIED UNSPECIFIED UNSPECIFIED Reuten, Raphael UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-126296
DOI:	10.1038/s41596-019-0225-8
Journal or Publication Title:	Nat. Protoc.
Volume:	14
Number:	12
Page Range:	S. 3395 - 3426
Date:	2019
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	1750-2799
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language PROTEOMIC ANALYSIS; PERFUSION; SCAFFOLDS; MULTIPLEX; ORGANS; BRAIN Multiple languages Biochemical Research Methods Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/12629