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.
This is a preview of subscription content
Subscribe to Nature+
Get immediate online access to the entire Nature family of 50+ journals
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Orkin, R. W. et al. A murine tumor producing a matrix of basement membrane. J. Exp. Med. 145, 204–220 (1977).
LeBleu, V. S., Macdonald, B. & Kalluri, R. Structure and function of basement membranes. Exp. Biol. Med. 232, 1121–1129 (2007).
Kubota, Y., Kleinman, H. K., Martin, G. R. & Lawley, T. J. Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J. Cell Biol. 107, 1589–1598 (1988).
Kleinman, H. K. et al. Basement membrane complexes with biological activity. Biochemistry 25, 312–318 (1986). This paper investigates the protein composition and biological activity of the basement-membrane extract from EHS mouse chondrosarcomas; this extract was later developed and commercialized as Matrigel.
Kleinman, H. K. & Martin, G. R. Matrigel: basement membrane matrix with biological activity. Semin. Cancer Biol. 15, 378–386 (2005).
Corning Incorporated Life Sciences. Corning Matrigel matrix. Frequently asked questions (Corning, 2019).
Hughes, C. S., Postovit, L. M. & Lajoie, G. A. Matrigel: a complex protein mixture required for optimal growth of cell culture. Proteomics 10, 1886–1890 (2010). A full proteomic analysis of Matrigel and GFR Matrigel, reporting their complex, ill-defined and variable composition.
Timpl, R. et al. Laminin — a glycoprotein from basement membranes. J. Biol. Chem. 254, 9933–9937 (1979).
Terranova, V. P., Aumailley, M., Sultan, L. H., Martin, G. R. & Kleinman, H. K. Regulation of cell attachment and cell number by fibronectin and laminin. J. Cell. Physiol. 127, 473–479 (1986).
Miyazaki, T. et al. Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells. Biochem. Biophys. Res. Commun. 375, 27–32 (2008).
Ponce, M. L. et al. Identification of endothelial cell binding sites on the laminin γ1 chain. Circ. Res. 84, 688–694 (1999).
Wang, K., Ji, L. & Hua, Z. Functional peptides from laminin-1 improve the cell adhesion capacity of recombinant mussel adhesive protein. Protein Pept. Lett. 24, 348–352 (2017).
Heaton, M. B. & Swanson, D. J. The influence of laminin on the initial differentiation of cultured neural tube neurons. J. Neurosci. Res. 19, 212–218 (1988).
Farrukh, A. et al. Bifunctional hydrogels containing the laminin motif IKVAV promote neurogenesis. Stem Cell Rep. 9, 1432–1440 (2017).
Ali, S., Saik, J. E., Gould, D. J., Dickinson, M. E. & West, J. L. Immobilization of cell-adhesive laminin peptides in degradable PEGDA hydrogels influences endothelial cell tubulogenesis. BioResearch Open Access 2, 241–249 (2013).
Engbring, J. A. & Kleinman, H. K. The basement membrane matrix in malignancy. J. Pathol. 200, 465–470 (2003).
Kikkawa, Y. et al. Laminin-111-derived peptides and cancer. Cell Adh. Migr. 7, 150–159 (2013).
Vukicevic, S. et al. Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular matrix components. Exp. Cell Res. 202, 1–8 (1992). This study identifies multiple active growth factors in Matrigel and suggests caution when interpreting cellular activity when cultured on Matrigel.
Talbot, N. C. & Caperna, T. J. Proteome array identification of bioactive soluble proteins/peptides in Matrigel: relevance to stem cell responses. Cytotechnology 67, 873–883 (2015).
Gillette, K. M., Forbes, K. & Sehgal, I. Detection of matrix metalloproteinases (MMP), tissue inhibitor of metalloproteinase-2, urokinase and plasminogen activator inhibitor-1 within Matrigel and growth factor-reduced Matrigel basement membrane. Tumori 89, 421–425 (2003).
Xu, C. et al. Feeder-free growth of undifferentiated human embryonic stem cells. Nat. Biotechnol. 19, 971–974 (2001).
Qian, L. & Saltzman, W. M. Improving the expansion and neuronal differentiation of mesenchymal stem cells through culture surface modification. Biomaterials 25, 1331–1337 (2004).
Lee, S.-W. et al. Optimization of Matrigel-based culture for expansion of neural stem cells. Anim. Cell Syst. 19, 175–180 (2015).
Laflamme, M. A. et al. Cardiomyocytes derived from human embryonic stem cells in pro-survival factors enhance function of infarcted rat hearts. Nat. Biotechnol. 25, 1015–1024 (2007).
Ponce, M. L. Tube formation: an in vitro Matrigel angiogenesis assay. Methods Mol. Biol. 467, 183–188 (2009).
Ponce, M. L. In vitro Matrigel angiogenesis assays. Methods Mol. Med. 46, 205–209 (2001).
Mondrinos, M. J. et al. Engineering three-dimensional pulmonary tissue constructs. Tissue Eng. 12, 717–728 (2006).
Li, Z. & Guan, J. Hydrogels for cardiac tissue engineering. Polymers 3, 740–761 (2011).
Lancaster, M. A. & Knoblich, J. A. Organogenesis in a dish: modeling development and disease using organoid technologies. Science 345, 1247125 (2014).
Lancaster, M. A. et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373–379 (2013).
Benton, G., Kleinman, H. K., George, J. & Arnaoutova, I. Multiple uses of basement membrane-like matrix (BME/Matrigel) in vitro and in vivo with cancer cells. Int. J. Cancer 128, 1751–1757 (2011).
Cruz-Acuña, R. & García, A. J. Synthetic hydrogels mimicking basement membrane matrices to promote cell-matrix interactions. Matrix Biol. 57–58, 324–333 (2017).
Polykandriotis, E., Arkudas, A., Horch, R. E., Kneser, U. & Mitchell, G. To Matrigel or not to Matrigel. Am. J. Pathol. 172, 1441–1442 (2008).
Kohen, N. T., Little, L. E. & Healy, K. E. Characterization of Matrigel interfaces during defined human embryonic stem cell culture. Biointerphases 4, 69–79 (2009).
Soofi, S. S., Last, J. A., Liliensiek, S. J., Nealey, P. F. & Murphy, C. J. The elastic modulus of Matrigel as determined by atomic force microscopy. J. Struct. Biol. 167, 216–219 (2009).
Dirami, G. et al. Identification of transferrin and inhibin-like proteins in Matrigel. In Vitro Cell. Dev. Biol. Anim. 31, 409–411 (1995).
Hansen, K. C. et al. An in-solution ultrasonication-assisted digestion method for improved extracellular matrix proteome coverage. Mol. Cell. Proteom. 8, 1648–1657 (2009).
Zaman, M. H. et al. Migration of tumor cells in 3D matrices is governed by matrix stiffness along with cell-matrix adhesion and proteolysis. Proc. Natl Acad. Sci. USA 103, 10889–10894 (2006).
Semler, E. J., Ranucci, C. S. & Moghe, P. V. Mechanochemical manipulation of hepatocyte aggregation can selectively induce or repress liver-specific function. Biotechnol. Bioeng. 69, 359–369 (2000).
Kane, K. I. W. et al. Determination of the rheological properties of Matrigel for optimum seeding conditions in microfluidic cell cultures. AIP Adv. 8, 125332 (2018).
Alcaraz, J. et al. Laminin and biomimetic extracellular elasticity enhance functional differentiation in mammary epithelia. EMBO J. 27, 2829–2838 (2008).
Reed, J., Walczak, W. J., Petzold, O. N. & Gimzewski, J. K. In situ mechanical interferometry of Matrigel films. Langmuir 25, 36–39 (2009).
Peterson, N. C. From bench to cageside: risk assessment for rodent pathogen contamination of cells and biologics. ILAR J. 49, 310–315 (2008).
Liu, H. et al. Removal of lactate dehydrogenase-elevating virus from human-in-mouse breast tumor xenografts by cell-sorting. J. Virol. Methods 173, 266–270 (2011).
Ammann, C. G., Messer, R. J., Peterson, K. E. & Hasenkrug, K. J. Lactate dehydrogenase-elevating virus induces systemic lymphocyte activation via TLR7-dependent IFNα responses by plasmacytoid dendritic cells. PLoS One 4, e6105 (2009).
Riley, V. et al. The LDH virus: an interfering biological contaminant. Science 200, 124–126 (1978).
Caliari, S. R. & Burdick, J. A. A practical guide to hydrogels for cell culture. Nat. Methods 13, 405–414 (2016).
Li, X., Sun, Q., Li, Q., Kawazoe, N. & Chen, G. Functional hydrogels with tunable structures and properties for tissue engineering applications. Front. Chem. 6, 499 (2018).
Fischer, R. S., Myers, K. A., Gardel, M. L. & Waterman, C. M. Stiffness-controlled three-dimensional extracellular matrices for high-resolution imaging of cell behavior. Nat. Protoc. 7, 2056–2066 (2012).
Tse, J. R. & Engler, A. J. Preparation of hydrogel substrates with tunable mechanical properties. Curr. Protoc. Cell Biol. 47, 10.16.1–10.16.16 (2010).
Pelham, R. J. Jr. & Wang, Y.-l. Cell locomotion and focal adhesions are regulated by substrate flexibility. Proc. Natl Acad. Sci. USA 94, 13661–13665 (1997).
Zustiak, S. P. & Leach, J. B. Hydrolytically degradable poly(ethylene glycol) hydrogel scaffolds with tunable degradation and mechanical properties. Biomacromolecules 11, 1348–1357 (2010).
Krsko, P. & Libera, M. Biointeractive hydrogels. Mater. Today 8, 36–44 (2005).
Zhu, J. Bioactive modification of poly(ethylene glycol) hydrogels for tissue engineering. Biomaterials 31, 4639–4656 (2010).
Lin, C.-C. & Anseth, K. S. PEG hydrogels for the controlled release of biomolecules in regenerative medicine. Pharm. Res. 26, 631–643 (2009).
Tibbitt, M. W. & Anseth, K. S. Hydrogels as extracellular matrix mimics for 3D cell culture. Biotechnol. Bioeng. 103, 655–663 (2009).
Fairbanks, B. D. et al. A versatile synthetic extracellular matrix mimic via thiol-norbornene photopolymerization. Adv. Mater. 21, 5005–5010 (2009).
Bryant, S. & Anseth, K. in Scaffolding in Tissue Engineering (eds Ma, P. X. & Elisseeff, J.) 71–90 (CRC, 2005).
Nguyen, K. T. & West, J. L. Photopolymerizable hydrogels for tissue engineering applications. Biomaterials 23, 4307–4314 (2002).
Nair, D. P. et al. The thiol-Michael addition click reaction: a powerful and widely used tool in materials chemistry. Chem. Mater. 26, 724–744 (2014).
Schense, J. C. & Hubbell, J. A. Cross-linking exogenous bifunctional peptides into fibrin gels with factor XIIIa. Bioconjug. Chem. 10, 75–81 (1999).
Ehrbar, M. et al. Biomolecular hydrogels formed and degraded via site-specific enzymatic reactions. Biomacromolecules 8, 3000–3007 (2007).
Bryant, S. J., Chowdhury, T. T., Lee, D. A., Bader, D. L. & Anseth, K. S. Crosslinking density influences chondrocyte metabolism in dynamically loaded photocrosslinked poly(ethylene glycol) hydrogels. Ann. Biomed. Eng. 32, 407–417 (2004).
Roberts, J. J. & Bryant, S. J. Comparison of photopolymerizable thiol-ene PEG and acrylate-based PEG hydrogels for cartilage development. Biomaterials 34, 9969–9979 (2013).
Burdick, J. A. & Anseth, K. S. Photoencapsulation of osteoblasts in injectable RGD-modified PEG hydrogels for bone tissue engineering. Biomaterials 23, 4315–4323 (2002).
Kharkar, P. M., Rehmann, M. S., Skeens, K. M., Maverakis, E. & Kloxin, A. M. Thiol–ene click hydrogels for therapeutic delivery. ACS Biomater. Sci. Eng. 2, 165–179 (2016).
Thomson, J. A. et al. Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147 (1998).
Avior, Y., Sagi, I. & Benvenisty, N. Pluripotent stem cells in disease modelling and drug discovery. Nat. Rev. Mol. Cell Biol. 17, 170–182 (2016).
Singh, V. K., Kalsan, M., Kumar, N., Saini, A. & Chandra, R. Induced pluripotent stem cells: applications in regenerative medicine, disease modeling, and drug discovery. Front. Cell Dev. Biol. 3, 2 (2015).
Shi, Y., Inoue, H., Wu, J. C. & Yamanaka, S. Induced pluripotent stem cell technology: a decade of progress. Nat. Rev. Drug. Discov. 16, 115–130 (2017).
Ortiz-Vitali, J. L. & Darabi, R. iPSCs as a platform for disease modeling, drug screening, and personalized therapy in muscular dystrophies. Cells 8, 20 (2019).
Hovatta, O. Derivation of human embryonic stem cell lines, towards clinical quality. Reprod. Fertil. Dev. 18, 823–828 (2006).
Qian, X., Villa-Diaz, L. G., Kumar, R., Lahann, J. & Krebsbach, P. H. Enhancement of the propagation of human embryonic stem cells by modifications in the gel architecture of PMEDSAH polymer coatings. Biomaterials 35, 9581–9590 (2014).
Nandivada, H. et al. Fabrication of synthetic polymer coatings and their use in feeder-free culture of human embryonic stem cells. Nat. Protoc. 6, 1037–1043 (2011).
Villa-Diaz, L. G. et al. Synthetic polymer coatings for long-term growth of human embryonic stem cells. Nat. Biotechnol. 28, 581–583 (2010). Along with reference 74, this was one of the first studies to develop a fully synthetic, chemically defined scaffold for long-term hESC culture and to directly compare the performance with that of Matrigel.
Brafman, D. A. et al. Long-term human pluripotent stem cell self-renewal on synthetic polymer surfaces. Biomaterials 31, 9135–9144 (2010).
Meng, Y. et al. Characterization of integrin engagement during defined human embryonic stem cell culture. FASEB J. 24, 1056–1065 (2009).
Rowland, T. J. et al. Roles of integrins in human induced pluripotent stem cell growth on Matrigel and vitronectin. Stem Cell Dev. 19, 1231–1240 (2010).
Mondal, G., Barui, S. & Chaudhuri, A. The relationship between the cyclic-RGDfK ligand and αvβ3 integrin receptor. Biomaterials 34, 6249–6260 (2013).
Lambshead, J. W. et al. Long-term maintenance of human pluripotent stem cells on cRGDfK-presenting synthetic surfaces. Sci. Rep. 8, 701 (2018).
Nguyen, E. H. et al. Versatile synthetic alternatives to Matrigel for vascular toxicity screening and stem cell expansion. Nat. Biomed. Eng. 1, 0096 (2017). This study uses a high-throughput screening method of synthetic scaffolds to determine a synthetic alternative to Matrigel, finding that matrix-induced effects caused by the biological function of Matrigel can affect toxicity screenings.
Hayman, E. G., Pierschbacher, M. D., Suzuki, S. & Ruoslahti, E. Vitronectin — a major cell attachment-promoting protein in fetal bovine serum. Exp. Cell Res. 160, 245–258 (1985).
Melkoumian, Z. et al. Synthetic peptide-acrylate surfaces for long-term self-renewal and cardiomyocyte differentiation of human embryonic stem cells. Nat. Biotechnol. 28, 606–610 (2010). An early report on tethering synthetic peptides to synthetic scaffolds that provides a direct comparison with Matrigel.
Deng, Y. et al. Long-term self-renewal of human pluripotent stem cells on peptide-decorated poly(OEGMA-co-HEMA) brushes under fully defined conditions. Acta Biomater. 9, 8840–8850 (2013).
Higuchi, A. et al. Long-term xeno-free culture of human pluripotent stem cells on hydrogels with optimal elasticity. Sci. Rep. 5, 18136 (2015).
Jin, S., Yao, H., Weber, J. L., Melkoumian, Z. K. & Ye, K. A synthetic, xeno-free peptide surface for expansion and directed differentiation of human induced pluripotent stem cells. PLoS One 7, e50880 (2012).
Yasuda, S. et al. Chemically defined and growth-factor-free culture system for the expansion and derivation of human pluripotent stem cells. Nat. Biomed. Eng. 2, 173–182 (2018).
Farach-Carson, M. C. & Carson, D. D. Perlecan — a multifunctional extracellular proteoglycan scaffold. Glycobiology 17, 897–905 (2007).
Furue, M. K. et al. Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium. Proc. Natl Acad. Sci. USA 105, 13409–13414 (2008).
Spivak-Kroizman, T. et al. Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation. Cell 79, 1015–1024 (1994).
Vlodavsky, I., Miao, H. Q., Medalion, B., Danagher, P. & Ron, D. Involvement of heparan sulfate and related molecules in sequestration and growth promoting activity of fibroblast growth factor. Cancer Metastasis Rev. 15, 177–186 (1996).
Chang, C.-W. et al. Engineering cell–material interfaces for long-term expansion of human pluripotent stem cells. Biomaterials 34, 912–921 (2013).
Klim, J. R., Li, L., Wrighton, P. J., Piekarczyk, M. S. & Kiessling, L. L. A defined glycosaminoglycan-binding substratum for human pluripotent stem cells. Nat. Methods 7, 989–994 (2010).
Musah, S. et al. Glycosaminoglycan-binding hydrogels enable mechanical control of human pluripotent stem cell self-renewal. ACS Nano 6, 10168–10177 (2012).
Gerecht, S. et al. Hyaluronic acid hydrogel for controlled self-renewal and differentiation of human embryonic stem cells. Proc. Natl Acad. Sci. USA 104, 11298–11303 (2007).
Lei, Y. & Schaffer, D. V. A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation. Proc. Natl Acad. Sci. USA 110, E5039–E5048 (2013).
Ovadia, E. M., Colby, D. W. & Kloxin, A. M. Designing well-defined photopolymerized synthetic matrices for three-dimensional culture and differentiation of induced pluripotent stem cells. Biomater. Sci. 6, 1358–1370 (2018).
Murphy, W. L., McDevitt, T. C. & Engler, A. J. Materials as stem cell regulators. Nat. Mater. 13, 547–557 (2014).
Folkman, J. & Moscona, A. Role of cell shape in growth control. Nature 273, 345–349 (1978).
Caiazzo, M. et al. Defined three-dimensional microenvironments boost induction of pluripotency. Nat. Mater. 15, 344–352 (2016).
Khalil, A. S., Xie, A. W. & Murphy, W. L. Context clues: the importance of stem cell–material interactions. ACS Chem. Biol. 9, 45–56 (2014).
Eve, D. J. The continued promise of stem cell therapy in regenerative medicine. Med. Sci. Monit. 17, RA292–RA305 (2011).
Helmy, K. Y., Patel, S. A., Silverio, K., Pliner, L. & Rameshwar, P. Stem cells and regenerative medicine: accomplishments to date and future promise. Ther. Deliv. 1, 693–705 (2010).
Hoffman, T., Khademhosseini, A. & Langer, R. Chasing the paradigm: clinical translation of 25 years of tissue engineering. Tissue Eng. Part A 25, 679–687 (2019).
Hwang, N. S., Varghese, S. & Elisseeff, J. Controlled differentiation of stem cells. Adv. Drug Deliv. Rev. 60, 199–214 (2008).
Burdick, J. A. & Vunjak-Novakovic, G. Engineered microenvironments for controlled stem cell differentiation. Tissue Eng. Part A 15, 205–219 (2009).
Lutolf, M. P., Gilbert, P. M. & Blau, H. M. Designing materials to direct stem-cell fate. Nature 462, 433–441 (2009).
Uriel, S. et al. Extraction and assembly of tissue-derived gels for cell culture and tissue engineering. Tissue Eng. Part C 15, 309–321 (2009).
Enemchukwu, N. O. et al. Synthetic matrices reveal contributions of ECM biophysical and biochemical properties to epithelial morphogenesis. J. Cell Biol. 212, 113–124 (2016).
Le, N. N. T., Zorn, S., Schmitt, S. K., Gopalan, P. & Murphy, W. L. Hydrogel arrays formed via differential wettability patterning enable combinatorial screening of stem cell behavior. Acta Biomater. 34, 93–103 (2016).
Koepsel, J. T., Brown, P. T., Loveland, S. G., Li, W.-J. & Murphy, W. L. Combinatorial screening of chemically defined human mesenchymal stem cell culture substrates. J. Mater. Chem. 22, 19474–19481 (2012).
Koutsopoulos, S. & Zhang, S. Long-term three-dimensional neural tissue cultures in functionalized self-assembling peptide hydrogels, Matrigel and Collagen I. Acta Biomater. 9, 5162–5169 (2013).
Zhang, J. et al. A genome-wide analysis of human pluripotent stem cell-derived endothelial cells in 2D or 3D culture. Stem Cell Rep. 8, 907–918 (2017).
Farhat, W. et al. Hydrogels for advanced stem cell therapies: a biomimetic materials approach for enhancing natural tissue function. IEEE Rev. Biomed. Eng. 12, 333–351 (2019).
Tsou, Y.-H., Khoneisser, J., Huang, P.-C. & Xu, X. Hydrogel as a bioactive material to regulate stem cell fate. Bioact. Mater. 1, 39–55 (2016).
Donnelly, H., Salmeron-Sanchez, M. & Dalby, M. J. Designing stem cell niches for differentiation and self-renewal. J. R. Soc. Interface 15, 20180388 (2018).
Vining, K. H. & Mooney, D. J. Mechanical forces direct stem cell behaviour in development and regeneration. Nat. Rev. Mol. Cell Biol. 18, 728–742 (2017).
Engler, A. J., Sen, S., Sweeney, H. L. & Discher, D. E. Matrix elasticity directs stem cell lineage specification. Cell 126, 677–689 (2006).
Slater, K., Partridge, J. & Nandivada, H. Tuning the elastic moduli of Corning Matrigel and collagen I 3D matrices by varying the protein concentration (Corning, 2018).
Gobaa, S. et al. Artificial niche microarrays for probing single stem cell fate in high throughput. Nat. Methods 8, 949–955 (2011).
Rape, A. D., Zibinsky, M., Murthy, N. & Kumar, S. A synthetic hydrogel for the high-throughput study of cell–ECM interactions. Nat. Commun. 6, 8129 (2015).
Nemir, S. & West, J. L. Synthetic materials in the study of cell response to substrate rigidity. Ann. Biomed. Eng. 38, 2–20 (2010).
Sill, T. J. & von Recum, H. A. Electrospinning: applications in drug delivery and tissue engineering. Biomaterials 29, 1989–2006 (2008).
Xu, C., Inai, R., Kotaki, M. & Ramakrishna, S. Electrospun nanofiber fabrication as synthetic extracellular matrix and its potential for vascular tissue engineering. Tissue Eng. 10, 1160–1168 (2004).
Rashidi, H., Yang, J. & M. Shakesheff, K. Surface engineering of synthetic polymer materials for tissue engineering and regenerative medicine applications. Biomater. Sci. 2, 1318–1331 (2014).
Yim, E. K. F., Pang, S. W. & Leong, K. W. Synthetic nanostructures inducing differentiation of human mesenchymal stem cells into neuronal lineage. Exp. Cell Res. 313, 1820–1829 (2007).
Zhu, W. et al. 3D printing of functional biomaterials for tissue engineering. Curr. Opin. Biotechnol. 40, 103–112 (2016).
Yamazoe, T. et al. A synthetic nanofibrillar matrix promotes in vitro hepatic differentiation of embryonic stem cells and induced pluripotent stem cells. J. Cell Sci. 126, 5391–5399 (2013).
Franzin, C. et al. Single-cell PCR analysis of murine embryonic stem cells cultured on different substrates highlights heterogeneous expression of stem cell markers. Biol. Cell 105, 549–560 (2013).
Highet, A. R., Zhang, V. J., Heinemann, G. K. & Roberts, C. T. Use of Matrigel in culture affects cell phenotype and gene expression in the first trimester trophoblast cell line HTR8/SVneo. Placenta 33, 586–588 (2012).
Sampaziotis, F. et al. Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells. Nat. Protoc. 12, 814–827 (2017).
Ranga, A. et al. Neural tube morphogenesis in synthetic 3D microenvironments. Proc. Natl Acad. Sci. USA 113, E6831–E6839 (2016).
Schneider, M. C. et al. Local heterogeneities improve matrix connectivity in degradable and photoclickable poly(ethylene glycol) hydrogels for applications in tissue engineering. ACS Biomater. Sci. Eng. 3, 2480–2492 (2017).
Chu, S. et al. Understanding the spatiotemporal degradation behavior of aggrecanase-sensitive poly(ethylene glycol) hydrogels for use in cartilage tissue engineering. Tissue Eng. Part A 23, 795–810 (2017).
Dolo, V. et al. Matrix-degrading proteinases are shed in membrane vesicles by ovarian cancer cells in vivo and in vitro. Clin. Exp. Metastasis 17, 131–140 (1999).
Balduyck, M. et al. Specific expression of matrix metalloproteinases 1, 3, 9 and 13 associated with invasiveness of breast cancer cells in vitro. Clin. Exp. Metastasis 18, 171–178 (2000).
Wong, A. P., Cortez, S. L. & Baricos, W. H. Role of plasmin and gelatinase in extracellular matrix degradation by cultured rat mesangial cells. Am. J. Physiol. 263, F1112–F1118 (1992).
Wolf, M. Influence of matrigel on biodistribution studies in cancer research. Pharmazie 63, 43–48 (2008).
Shen, D., Wen, R., Tuo, J., Bojanowski, C. M. & Chan, C.-C. Exacerbation of retinal degeneration and choroidal neovascularization induced by subretinal injection of Matrigel in CCL2/MCP-1-deficient mice. Ophthalmic Res. 38, 71–73 (2006).
Kano, M. R. et al. VEGF-A and FGF-2 synergistically promote neoangiogenesis through enhancement of endogenous PDGF-B-PDGFRbeta signaling. J. Cell Sci. 118, 3759–3768 (2005).
Lee, J. H. Injectable hydrogels delivering therapeutic agents for disease treatment and tissue engineering. Biomater. Res. 22, 27 (2018).
Yu, L. & Ding, J. Injectable hydrogels as unique biomedical materials. Chem. Soc. Rev. 37, 1473–1481 (2008).
Kretlow, J. D., Klouda, L. & Mikos, A. G. Injectable matrices and scaffolds for drug delivery in tissue engineering. Adv. Drug Deliv. Rev. 59, 263–273 (2007).
Pascual-Garrido, C. et al. Current and novel injectable hydrogels to treat focal chondral lesions: properties and applicability. J. Orthop. Res. 36, 64–75 (2018).
Kharkar, P. M., Kiick, K. L. & Kloxin, A. M. Designing degradable hydrogels for orthogonal control of cell microenvironments. Chem. Soc. Rev. 42, 7335–7372 (2013).
Han, W. M. et al. Synthetic matrix enhances transplanted satellite cell engraftment in dystrophic and aged skeletal muscle with comorbid trauma. Sci. Adv. 4, eaar4008 (2018).
Fernandes, S., Kuklok, S., McGonigle, J., Reinecke, H. & Murry, C. E. Synthetic matrices to serve as niches for muscle cell transplantation. Cells Tissues Organs 195, 48–59 (2012).
Nagahama, K. et al. Nanocomposite injectable gels capable of self-replenishing regenerative extracellular microenvironments for in vivo tissue engineering. Biomater. Sci. 6, 550–561 (2018).
Clevers, H. Modeling development and disease with organoids. Cell 165, 1586–1597 (2016).
Camp, J. G. et al. Human cerebral organoids recapitulate gene expression programs of fetal neocortex development. Proc. Natl Acad. Sci. USA 112, 15672–15677 (2015).
Murrow, L. M., Weber, R. J. & Gartner, Z. J. Dissecting the stem cell niche with organoid models: an engineering-based approach. Development 144, 998–1007 (2017).
Astashkina, A. I., Mann, B. K., Prestwich, G. D. & Grainger, D. W. A 3-D organoid kidney culture model engineered for high-throughput nephrotoxicity assays. Biomaterials 33, 4700–4711 (2012).
Takebe, T. et al. Vascularized and functional human liver from an iPSC-derived organ bud transplant. Nature 499, 481–484 (2013).
Eiraku, M. et al. Self-organizing optic-cup morphogenesis in three-dimensional culture. Nature 472, 51–56 (2011).
Dye, B. R. et al. In vitro generation of human pluripotent stem cell derived lung organoids. eLife 4, e05098 (2015).
Cruz-Acuña, R. et al. Synthetic hydrogels for human intestinal organoid generation and colonic wound repair. Nat. Cell Biol. 19, 1326–1335 (2017). The authors present a protocol to generate human intestinal and lung organoids using a fully synthetic, chemically defined PEG hydrogel scaffold.
Chua, C. W. et al. Single luminal epithelial progenitors can generate prostate organoids in culture. Nat. Cell Biol. 16, 951–961 (2014).
Ardalani, H. et al. 3-D culture and endothelial cells improve maturity of human pluripotent stem cell-derived hepatocytes. Acta Biomater. 95, 371–381 (2019).
Ramachandran, S. D. et al. In vitro generation of functional liver organoid-like structures using adult human cells. PLoS One 10, e0139345 (2015).
Spence, J. R. et al. Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro. Nature 470, 105–109 (2011).
Cruz-Acuña, R. et al. PEG-4MAL hydrogels for human organoid generation, culture, and in vivo delivery. Nat. Protoc. 13, 2102 (2018).
Gjorevski, N. et al. Designer matrices for intestinal stem cell and organoid culture. Nature 539, 560–564 (2016).
Gjorevski, N. & Lutolf, M. P. Synthesis and characterization of well-defined hydrogel matrices and their application to intestinal stem cell and organoid culture. Nat. Protoc. 12, 2263–2274 (2017).
Fatehullah, A., Tan, S. H. & Barker, N. Organoids as an in vitro model of human development and disease. Nat. Cell Biol. 18, 246–254 (2016).
Bray, L. J. et al. A three-dimensional ex vivo tri-culture model mimics cell-cell interactions between acute myeloid leukemia and the vascular niche. Haematologica 102, 1215–1226 (2017).
Papadimitriou, C. et al. 3D culture method for Alzheimer’s disease modeling reveals interleukin-4 rescues Aβ42-induced loss of human neural stem cell plasticity. Dev. Cell 46, 85–101.e8 (2018).
Nowak, M., Freudenberg, U., Tsurkan, M. V., Werner, C. & Levental, K. R. Modular GAG-matrices to promote mammary epithelial morphogenesis in vitro. Biomaterials 112, 20–30 (2017).
Weber, H. M., Tsurkan, M. V., Magno, V., Freudenberg, U. & Werner, C. Heparin-based hydrogels induce human renal tubulogenesis in vitro. Acta Biomater. 57, 59–69 (2017).
Livingston, M. K. et al. Evaluation of PEG-based hydrogel influence on estrogen-receptor-driven responses in MCF7 breast cancer cells. ACS Biomater. Sci. Eng. 5, 6089–6098 (2019).
Edmondson, R., Adcock, A. F. & Yang, L. Influence of matrices on 3D-cultured prostate cancer cells’ drug response and expression of drug-action associated proteins. PLoS One 11, e0158116 (2016).
Collier, J. H. & Segura, T. Evolving the use of peptides as biomaterials components. Biomaterials 32, 4198–4204 (2011).
Hosoyama, K., Lazurko, C., Muñoz, M., McTiernan, C. D. & Alarcon, E. I. Peptide-based functional biomaterials for soft-tissue repair. Front. Bioeng. Biotechnol. 7, 205 (2019).
Xie, A. W. & Murphy, W. L. Engineered biomaterials to mitigate growth factor cost in cell biomanufacturing. Curr. Opin. Biomed. Eng. 10, 1–10 (2019).
Heidariyan, Z. et al. Efficient and cost-effective generation of hepatocyte-like cells through microparticle-mediated delivery of growth factors in a 3D culture of human pluripotent stem cells. Biomaterials 159, 174–188 (2018).
Bratt-Leal, A. M., Nguyen, A. H., Hammersmith, K. A., Singh, A. & McDevitt, T. C. A microparticle approach to morphogen delivery within pluripotent stem cell aggregates. Biomaterials 34, 7227–7235 (2013).
Alberti, K. et al. Functional immobilization of signaling proteins enables control of stem cell fate. Nat. Methods 5, 645–650 (2008).
Platt, M. O. et al. Sustained epidermal growth factor receptor levels and activation by tethered ligand binding enhances osteogenic differentiation of multi-potent marrow stromal cells. J. Cell. Physiol. 221, 306–317 (2009).
Yu, X. et al. Nanostructured mineral coatings stabilize proteins for therapeutic delivery. Adv. Mater. 29, 1701255 (2017).
Khalil, A. S., Xie, A. W., Johnson, H. J. & Murphy, W. L. Sustained release and protein stabilization reduce the growth factor dosage required for human pluripotent stem cell expansion. Biomaterials 248, 120007 (2020).
Belair, D. G., Le, N. N. & Murphy, W. L. Design of growth factor sequestering biomaterials. Chem. Commun. 50, 15651–15668 (2014).
Belair, D. G. & Murphy, W. L. Specific VEGF sequestering to biomaterials: influence of serum stability. Acta Biomater. 9, 8823–8831 (2013).
Yan, H. J. et al. Synthetic design of growth factor sequestering extracellular matrix mimetic hydrogel for promoting in vivo bone formation. Biomaterials 161, 190–202 (2018).
Jha, A. K. et al. Enhanced survival and engraftment of transplanted stem cells using growth factor sequestering hydrogels. Biomaterials 47, 1–12 (2015).
Chen, K. G., Mallon, B. S., McKay, R. D. G. & Robey, P. G. Human pluripotent stem cell culture: considerations for maintenance, expansion, and therapeutics. Cell Stem Cell 14, 13–26 (2014).
Julavijitphong, S. et al. A xeno-free culture method that enhances Wharton’s jelly mesenchymal stromal cell culture efficiency over traditional animal serum-supplemented cultures. Cytotherapy 16, 683–691 (2014).
Thirumala, S., Goebel, W. S. & Woods, E. J. Manufacturing and banking of mesenchymal stem cells. Expert Opin. Biol. Ther. 13, 673–691 (2013).
Halme, D. G. & Kessler, D. A. FDA regulation of stem-cell-based therapies. N. Engl. J. Med. 355, 1730–1735 (2006).
Xiao, J., Yang, D., Li, Q., Tian, W. & Guo, W. The establishment of a chemically defined serum-free culture system for human dental pulp stem cells. Stem Cell Res. Ther. 9, 191 (2018).
Hirata, T. M. et al. Expression of multiple stem cell markers in dental pulp cells cultured in serum-free media. J. Endod. 36, 1139–1144 (2010).
Chen, G. et al. Chemically defined conditions for human iPSC derivation and culture. Nat. Methods 8, 424–429 (2011).
Beers, J. et al. A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture. Sci. Rep. 5, 11319 (2015).
Xie, A. W. et al. Controlled self-assembly of stem cell aggregates instructs pluripotency and lineage bias. Sci. Rep. 7, 14070 (2017).
Leong, M. F. et al. Electrospun polystyrene scaffolds as a synthetic substrate for xeno-free expansion and differentiation of human induced pluripotent stem cells. Acta Biomater. 46, 266–277 (2016).
This research was funded by the US Environmental Protection Agency (STAR grant no. 83573701), the US National Institutes of Health (award nos. 1U01TR002383, R01HL093282 and 1R01NS109427) and the US National Science Foundation (award nos. EEC1648035 and DMR 170179). E.A.A. acknowledges funding by the US National Institutes of Health (T32HL110853).
W.L.M. is a co-founder and shareholder of Stem Pharm, Inc. E.A.A. declares no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Aisenbrey, E.A., Murphy, W.L. Synthetic alternatives to Matrigel. Nat Rev Mater 5, 539–551 (2020). https://doi.org/10.1038/s41578-020-0199-8
Journal of Biomedical Science (2022)
Communications Biology (2022)
Apical-out airway organoids as a platform for studying viral infections and screening for antiviral drugs
Scientific Reports (2022)
Tissue extracellular matrix hydrogels as alternatives to Matrigel for culturing gastrointestinal organoids
Nature Communications (2022)
Nature Materials (2022)