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Decellularization and antibody staining of mouse tissues to map native extracellular matrix structures in 3D

Mayorca-Guiliani, Alejandro E ; Willacy, Oliver ; Madsen, Chris D LU ; Rafaeva, Maria ; Elisabeth Heumüller, Stefanie ; Bock, Felix ; Sengle, Gerhard ; Koch, Manuel ; Imhof, Thomas and Zaucke, Frank , et al. (2019) In Nature Protocols 14. p.3395-3425
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... (More)

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.

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publishing date
type
Contribution to journal
publication status
published
subject
in
Nature Protocols
volume
14
pages
3395 - 3425
publisher
Nature Publishing Group
external identifiers
  • scopus:85075132942
  • pmid:31705125
ISSN
1750-2799
DOI
10.1038/s41596-019-0225-8
language
English
LU publication?
yes
id
058c31f0-2d9e-4676-a1eb-7f0c90ce6ea5
date added to LUP
2019-11-15 11:24:51
date last changed
2020-10-27 02:57:45
@article{058c31f0-2d9e-4676-a1eb-7f0c90ce6ea5,
  abstract     = {<p>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.</p>},
  author       = {Mayorca-Guiliani, Alejandro E and Willacy, Oliver and Madsen, Chris D and Rafaeva, Maria and Elisabeth Heumüller, Stefanie and Bock, Felix and Sengle, Gerhard and Koch, Manuel and Imhof, Thomas and Zaucke, Frank and Wagener, Raimund and Sasaki, Takako and Erler, Janine T and Reuten, Raphael},
  issn         = {1750-2799},
  language     = {eng},
  pages        = {3395--3425},
  publisher    = {Nature Publishing Group},
  series       = {Nature Protocols},
  title        = {Decellularization and antibody staining of mouse tissues to map native extracellular matrix structures in 3D},
  url          = {http://dx.doi.org/10.1038/s41596-019-0225-8},
  doi          = {10.1038/s41596-019-0225-8},
  volume       = {14},
  year         = {2019},
}