Decellularization and antibody staining of mouse tissues to map native extracellular matrix structures in 3D
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.
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SDC is more biocompatible compared to SDS.
“SDC is also more biocompatible compared to SDS [48, 91].”
From Immunogenicity of decellularized extracellular matrix scaffolds: a bottleneck in tissue engineering and regenerative medicine (2023) · cites this paper as [91]
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- Authors
- Mayorca-Guiliani, A. E., Willacy, O., Madsen, C. D., Rafaeva, M., Elisabeth Heumüller, S., Bock, F., Sengle, G., Koch, M., Imhof, T., Zaucke, F., Wagener, R., Sasaki, T., Erler, J. T., & Reuten, R.
- Affiliations
- University of Copenhagen (DK)
- Lund University (SE)
- University of Cologne (DE)
- Orthopaedic University Hospital Friedrichsheim (DE)
- Oita University (JP)
- Corresponding author
- Erler, Janine T.
- Reuten, Raphael
- ORCID
- Venue
- Nature Protocols
- Published
- 2019-11-08
- Volume
- 14
- Issue
- 12
- Pages
- 3395-3425
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2983262330
- Cited by
- 79
- References
- 5
- Citations by year
- 2026: 5 · 2025: 8 · 2024: 11 · 2023: 14 · 2022: 14 · 2021: 17 · 2020: 10
- FWCI
- 9.88
- Type
- journal article
- Language
- en
- Update notices
- correction (2020-03-16)
- Open access
- closed
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Kölner Universitäts PublikationsServer (Universität zu Köln) (publishedVersion)
- Lund University Publications (Lund University) (submittedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (submittedVersion)
- Funders
- Lundbeckfonden (R286-2018-621)
- Novo Nordisk Fonden (NNF12OC1015992)
- Topics
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Keywords
- extracellular matrix, decellularization, antibody staining, perfusion decellularization, mouse tissues, tissue engineering scaffolds
- MeSH terms
- Animals, Antibodies, Extracellular Matrix, Mice, Inbred BALB C, Mice, Inbred C57BL, Organ Specificity, Perfusion, Staining and Labeling, Imaging, Three-Dimensional, Tissue Engineering, Mice, Tissue Scaffolds
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