Abstract
Altered tissue mechanical properties have been implicated in many key physiological and pathological processes. Hydrogel-based materials systems, made with native extracellular matrix (ECM) proteins, nonnative biopolymers, or synthetic polymers are often used to study these processes in vitro in 3D cell culture experiments. However, each of these materials systems present major limitations when used in mechanobiological studies. While native ECM-based hydrogels may enable good recapitulation of physiological behavior, the mechanics of these hydrogels are often manipulated by increasing or decreasing the protein concentration. This manipulation changes cell adhesion ligand density, thereby altering cell signaling. Alternatively, synthetic polymer-based hydrogels and nonnative biopolymer-based hydrogels can be mechanically tuned and engineered to present cell adhesion peptide motifs, but still may not fully promote physiologically relevant behavior. Here, we combine the advantages of native ECM proteins and nonnative biopolymers in interpenetrating network (IPN) hydrogels consisting of rBM matrix, which contains ligands native to epithelial basement membrane, and alginate, an inert biopolymer derived from seaweed. The following protocol details the generation of IPNs for mechanical testing or for 3D cell culture. This biomaterial system offers the ability to tune the stiffness of the 3D microenvironment without altering cell adhesion ligand concentration or pore size.
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References
Eyckmans J, Boudou T, Yu X et al (2011) A hitchhiker’s guide to mechanobiology. Dev Cell 21:35–47
Pedersen JA, Swartz MA (2005) Mechanobiology in the third dimension. Ann Biomed Eng 33:1469–1490
Tibbitt MW, Anseth KS (2009) Hydrogels as extracellular matrix mimics for 3D cell culture. Biotechnol Bioeng 103:655–663
Cukierman E, Pankov R, Stevens DR et al (2001) Taking cell-matrix adhesions to the third dimension. Science 294:1708–1712
Chaudhuri O, Koshy S, Branco da Cunha C et al (2014) Extracellular matrix stiffness and composition jointly regulate the induction of malignant phenotypes in mammary epithelium. Nat Mater 13:970–978
Mammoto A, Connor KM, Mammoto T et al (2009) A mechanosensitive transcriptional mechanism that controls angiogenesis. Nature 457:1103–1108
Boyd-White J, Williams JC (1996) Effect of cross-linking on matrix permeability: a model for AGE-modified basement membranes. Diabetes 45:348–353
Kleinman HK, Martin GR (2005) Matrigel : basement membrane matrix with biological activity. Semin Cancer Biol 15:378–386
Yurchenco PD, Schittny JC (1990) Molecular architecture of basement membranes. FASEB J 4:1577–1590
Rowley JA, Madlambayan G, Mooney DJ (1999) Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials 20:45–53
Rehm BHA (2009) Alginates: biology and applications. In: Steinbüchel A (ed) Microbiology monographs, vol 13. Springer, Heidelberg
Huebsch N, Arany PR, Mao AS et al (2010) Harnessing traction-mediated manipulation of the cell/matrix interface to control stem-cell fate. Nat Mater 9:518–526
Kong HJ, Kim CJ, Huebsch N et al (2007) Noninvasive probing of the spatial organization of polymer chains in hydrogels using fluorescence resonance energy transfer (FRET). J Am Chem Soc 129:4518–4519
Acknowledgements
This work was supported by the National Science Foundation Graduate Research Fellowship to Katrina Wisdom.
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Wisdom, K., Chaudhuri, O. (2017). 3D Cell Culture in Interpenetrating Networks of Alginate and rBM Matrix. In: Koledova, Z. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 1612. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7021-6_3
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DOI: https://doi.org/10.1007/978-1-4939-7021-6_3
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7019-3
Online ISBN: 978-1-4939-7021-6
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