Matrigel: A complex protein mixture required for optimal growth of cell culture
Abstract
Numerous cell types require a surface for attachment to grow and proliferate. Certain cells, particularly primary and stem cells, necessitate the use of specialized growth matrices along with specific culture media conditions to maintain the cells in an undifferentiated state. A gelatinous protein mixture derived from mouse tumor cells and commercialized as Matrigel is commonly used as a basement membrane matrix for stem cells because it retains the stem cells in an undifferentiated state. However, Matrigel is not a well‐defined matrix, and therefore can produce a source of variability in experimental results. In this study, we present an in‐depth proteomic analysis of Matrigel using a dynamic iterative exclusion method coupled with fractionation protocols that involve ammonium sulfate precipitation, size exclusion chromatography, and one‐dimensional SDS‐PAGE. The ability to identify the low mass and abundance components of Matrigel illustrates the utility of this method for the analysis of the extracellular matrix, as well as the complexity of the matrix itself.
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Matrigel is a mixture of the basement membrane proteins collagen type IV, laminin, entactin and heparan sulphate proteoglycane.
“Matrigel is a mixture of the BM proteins collagen type IV, laminin, entactin and heparan sulphate proteoglycane (Benton et al., 2014[4]; Hughes et al., 2010[19]).”
From Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-matrigel matrices (2023) · cites this paper as Hughes et al., 2010[19]
Culturing hepatocytes on either collagen type I or Matrigel alone does not sufficiently reflect the ECM composition of the liver.
“Culturing hepatocytes on either collagen type I or Matrigel alone, does not sufficiently reflect the ECM composition of the liver (Benton et al., 2014[4]; Hughes et al., 2010[19]; Martinez-Hernandez and Amenta, 1993[26]).”
From Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-matrigel matrices (2023) · cites this paper as Hughes et al., 2010[19]
The liver's ECM is much more complex than the most widely applied matrices for hepatocyte culture, namely collagen type I and Matrigel.
“The liver's ECM is much more complex than the most widely applied matrices for hepatocyte culture, namely collagen type I and Matrigel.”
From Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-matrigel matrices (2023) · cites this paper as Hughes et al., 2010[19]
Matrigel contains several hepatic ECM components, like collagen type IV, laminin and heparan sulfate proteoglycane.
“the basement membrane extract Matrigel contains several of these hepatic ECM components, like collagen type IV, laminin and heparan sulfate proteoglycane (Hughes et al., 2010[19]).”
From Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-matrigel matrices (2023) · cites this paper as Hughes et al., 2010[19]
The batch-to-batch and within-batch variability of Matrigel and other tumor- or tissue-derived ECM preparations hinders standardization of 3D and organoid cultures.
“For instance, Matrigel and other tumor- or tissue-derived ECM preparations exhibit pronounced batch-to-batch and even within-batch variability in biochemical and mechanical properties, hindering standardization of 3D and organoid cultures (Hughes et al., 2010; Aisenbrey and Murphy, 2020).”
From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Hughes et al., 2010
Matrigel and other tumor- or tissue-derived ECM preparations exhibit pronounced batch-to-batch and even within-batch variability in biochemical and mechanical properties.
“For instance, Matrigel and other tumor- or tissue-derived ECM preparations exhibit pronounced batch-to-batch and even within-batch variability in biochemical and mechanical properties, hindering standardization of 3D and organoid cultures (Hughes et al., 2010; Aisenbrey and Murphy, 2020).”
From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Hughes et al., 2010
Animal-derived matrices introduce significant limitations, including batch-to-batch compositional and mechanical variability and undefined growth factor content, which can confound lineage commitment and reproducibility.
“Despite their functional advantages, animal-derived matrices introduce significant limitations, including batch-to-batch compositional and mechanical variability and undefined growth factor content, which can confound lineage commitment and reproducibility (Hughes et al., 2010).”
From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Hughes et al., 2010
Matrigel is complex and poorly defined.
“Matrigel is complex and poorly defined.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [44]
Proteomic analysis shows that Matrigel contains more than 1800 unique proteins.
“Despite its versatility and affordability, Matrigel is extremely complex; proteomic analysis shows that it contains more than 1800 unique proteins44.”
From Towards organoid culture without Matrigel (2021) · cites this paper as [44]
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- Authors
- Hughes, C. S., Postovit, L. M., & Lajoie, G. A.
- Affiliations
- Western University (CA)
- Corresponding author
- Lajoie, Gilles A.
- Venue
- PROTEOMICS
- Publisher
- Wiley
- Published
- 2010-04-28
- Volume
- 10
- Issue
- 9
- Pages
- 1886-1890
- ISSN
- 1615-9853 (ISSN-L)
- 1615-9861
- OpenAlex
- W1971163936
- Cited by
- 1,534
- References
- 12
- Citations by year
- 2026: 109 · 2025: 113 · 2024: 130 · 2023: 122 · 2022: 155 · 2021: 177 · 2020: 164 · 2019: 99 · 2018: 82 · 2017: 94 · 2016: 83 · 2015: 61 · 2014: 61 · 2013: 36 · 2012: 29
- FWCI
- 12.04
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- PROTEOMICS (publishedVersion)
- PubMed (publishedVersion)
- Scholarship@Western (Western University) (submittedVersion)
- Funders
- NSERC Canada Graduate Scholarship doctoral award
- National Science and Engineering Council
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Health Sciences › Medicine › Rheumatology › Bone and Dental Protein Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Keywords
- Matrigel, extracellular matrix, proteomics, protein fractionation, ammonium sulfate precipitation, size exclusion chromatography, stem cell culture, primary cell culture, cell attachment, undifferentiated state, matrix heterogeneity
- MeSH terms
- Collagen, Drug Combinations, Laminin, Proteoglycans, Proteomics
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