Synthetic alternatives to Matrigel
Open accessAbstract
Matrigel, a basement-membrane matrix extracted from Engelbreth-Holm-Swarm mouse sarcomas, has been used for more than four decades for a myriad of cell culture applications. However, Matrigel is limited in its applicability to cellular biology, therapeutic cell manufacturing and drug discovery owing to its complex, ill-defined and variable composition. Variations in the mechanical and biochemical properties within a single batch of Matrigel - and between batches - have led to uncertainty in cell culture experiments and a lack of reproducibility. Moreover, Matrigel is not conducive to physical or biochemical manipulation, making it difficult to fine-tune the matrix to promote intended cell behaviours and achieve specific biological outcomes. Recent advances in synthetic scaffolds have led to the development of xenogenic-free, chemically defined, highly tunable and reproducible alternatives. In this Review, we assess the applications of Matrigel in cell culture, regenerative medicine and organoid assembly, detailing the limitations of Matrigel and highlighting synthetic scaffold alternatives that have shown equivalent or superior results. Additionally, we discuss the hurdles that are limiting a full transition from Matrigel to synthetic scaffolds and provide a brief perspective on the future directions of synthetic scaffolds for cell culture applications.
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Animal-derived reagents and human blood-derived products are frequently associated with undefined or incompletely characterized composition and batch-to-batch variability.
“These animal-derived reagents, and also human blood-derived products, are frequently associated with undefined or incompletely characterized composition and batch-to-batch variability, which can compromise reproducibility and limit translational relevance (Aisenbrey and Murphy, 2020; Cassotta et al., 2022; Nezvedova et al., 2025; Wolff and Hendrix, 2025; Akuta et al., 2026).”
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 Aisenbrey and Murphy, 2020
The 'Limitations of Matrigel' section describes animal-derived Matrigel as having a complex, ill-defined, variable composition and says differences between batches undermine reproducibility. It does not establish the same composition and batch-variability claim for human blood-derived products; its mention of serum-containing media does not identify such products or make that claim about them. (LLM, conf=0.88)
Experimental — accuracy not yet measuredThe undefined or incompletely characterized composition and batch-to-batch variability of animal-derived reagents and human blood-derived products can compromise reproducibility and limit translational relevance.
“These animal-derived reagents, and also human blood-derived products, are frequently associated with undefined or incompletely characterized composition and batch-to-batch variability, which can compromise reproducibility and limit translational relevance (Aisenbrey and Murphy, 2020; Cassotta et al., 2022; Nezvedova et al., 2025; Wolff and Hendrix, 2025; Akuta et al., 2026).”
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 Aisenbrey and Murphy, 2020
The ‘Limitations of Matrigel’ section says this animal-derived matrix has an ill-defined, variable composition that undermines reproducibility and limits its applicability to therapeutic cell manufacturing. The cited passages do not establish the same composition and batch-variability concerns for human blood-derived products; their mention of serum-containing media does not identify it as human-derived. (LLM, conf=0.87)
Experimental — accuracy not yet measuredSingle glycoproteins such as laminin, fibronectin, and vitronectin promote integrin-mediated adhesion.
“including the use of single glycoproteins such as laminin, fibronectin, and vitronectin, which promote integrin-mediated adhesion (Hayashi and Furue, 2016; Aisenbrey and Murphy, 2020; Kawase and Nakatsuji, 2023).”
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 Aisenbrey and Murphy, 2020
The Introduction describes adhesion sites on laminin, and the Pluripotent stem cell culture section explains that a fibronectin-derived peptide binds integrins and promotes adhesion. The review also discusses peptides derived from vitronectin, but these passages do not establish that each of the three whole glycoproteins, used singly, promotes integrin-mediated adhesion. (LLM, conf=0.82)
Experimental — accuracy not yet measuredMatrigel has an ill-defined chemistry and composition, and major lot-to-lot variation, and thus offers poor control over ECM composition.
“Although Matrigel provides important biochemical cues, it has an ill-defined chemistry and composition, and major lot-to-lot variation [18,19], and thus offers a poor control over ECM composition [20].”
Blood contamination, especially erythrocytes, can affect organoid generation and matrix stability.
“Blood contamination, particularly erythrocytes, can also affect organoid generation and matrix stability therefore standard approaches to eliminate these through lysis are typically used60.”
From Organoids (2022) · cites this paper as [60]
The Organoid assembly section discusses how Matrigel heterogeneity affects organoid formation, and Limitations of Matrigel discusses xenogenic and viral contaminants. Neither passage addresses blood contamination, erythrocytes, their effects on matrix stability, or removal by lysis; searches for erythrocytes and red blood cell lysis found no such passage. Page numbers were not available in our copy. (LLM, conf=0.72)
Experimental — accuracy not yet measured
Synthetic alternatives to Matrigel have been described.
“Synthetic alternatives to Matrigel have been described [26].”
From A worldwide survey on the use of animal‐derived materials and reagents in scientific experimentation (2022) · cites this paper as [26]
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 Aisenbrey and Murphy, 2020
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 Aisenbrey and Murphy, 2020
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Flagged first, then by citations
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Cited by
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“Furthermore, other ECM-derived matrices, such as Matrigel and decellularized ECM, are widely used in cell culture, in tissue engineering, and even as clinical therapies; however, their batch variability and animal sourcing are limiting (13).”
BackgroundExtracellular matrix requirements for gastrointestinal organoid cultures
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“Some synthetic, animal-free alternatives are now available (Aisenbrey and Murphy 2020), and the field of generating them is highly dynamic.”
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- Authors
- Aisenbrey, E. A., & Murphy, W. L.
- Affiliations
- University of Wisconsin–Madison (US)
- Corresponding author
- Murphy, William L.
- Venue
- Nature Reviews Materials
- Published
- 2020-05-27
- Volume
- 5
- Issue
- 7
- Pages
- 539-551
- ISSN
- 2058-8437 (ISSN-L)
- PMC
- PMC7500703
- OpenAlex
- W3028768728
- Cited by
- 1,016
- References
- 193
- Citations by year
- 2026: 167 · 2025: 211 · 2024: 176 · 2023: 170 · 2022: 155 · 2021: 117 · 2020: 19 · 2012: 1
- FWCI
- 13.46
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Nature Reviews Materials (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health (U01TR002383)
- National Heart, Lung, and Blood Institute (T32HL110853, R01HL093282)
- National Institute of Neurological Disorders and Stroke (R01NS109427)
- National Center for Advancing Translational Sciences
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Hydrogels: synthesis, properties, applications
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- Matrigel, synthetic scaffolds, batch-to-batch variation, stem cell culture, regenerative medicine, extracellular matrix
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