Abstract
The biggest challenge of designing and implementing an in vitro study is developing a microenvironment that most closely represents the interactions observed in vivo. Decellularization of tissues and organs has been shown to be an effective method for the removal of potentially immunogenic constituents while preserving essential growth factors and extracellular matrix (ECM ) proteins necessary for proper cell function. Enzymatic digestion of decellularized tissues allows these tissue-specific components to be reconstituted into bioactive hydrogels through a physical crosslinking of collagen. In the following protocol, we describe unique decellularization methods for both dermis and urinary bladder matrix (UBM) derived from porcine tissues. We then provide details for hydrogel formation and subsequent three-dimensional (3D) culture of two cell types: NIH 3T3 fibroblasts and C2C12 myoblasts .
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Koochekpour S, Merzak A, Pilkington GJ (1995) Extracellular matrix proteins inhibit proliferation, upregulate migration and induce morphological changes in human glioma cell lines. Eur J Cancer 31A(3):375–380
Williams CM, Engler AJ, Slone RD et al (2008) Fibronectin expression modulates mammary epithelial cell proliferation during acinar differentiation. Cancer Res 68(9):3185–3192
Bosnakovski D, Mizuno M, Kim G et al (2006) Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: influence of collagen type II extracellular matrix on MSC chondrogenesis. Biotechnol Bioeng 93(6):1152–1163
Chastain SR, Kundu AK, Dhar S et al (2006) Adhesion of mesenchymal stem cells to polymer scaffolds occurs via distinct ECM ligands and controls their osteogenic differentiation. J Biomed Mater Res A 78a(1):73–85
Simon-Assmann P, Kedinger M, De Arcangelis A et al (1995) Extracellular matrix components in intestinal development. Experientia 51(9–10):883–900
DeQuach JA, Mezzano V, Miglani A et al (2010) Simple and high yielding method for preparing tissue specific extracellular matrix coatings for cell culture. PLoS One 5(9):e13039
Lutolf MP, Hubbell JA (2005) Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol 23(1):47–55
Uriel S, Labay E, Francis-Sedlak M et al (2009) Extraction and assembly of tissue-derived gels for cell culture and tissue engineering. Tissue Eng Part C Methods 15(3):309–321
Stern MM, Myers RL, Hammam N et al (2009) The influence of extracellular matrix derived from skeletal muscle tissue on the proliferation and differentiation of myogenic progenitor cells ex vivo. Biomaterials 30(12):2393–2399
Zhang Y, He Y, Bharadwaj S et al (2009) Tissue-specific extracellular matrix coatings for the promotion of cell proliferation and maintenance of cell phenotype. Biomaterials 30(23–24):4021–4028
Young DA, Ibrahim DO, Hu D et al (2011) Injectable hydrogel scaffold from decellularized human lipoaspirate. Acta Biomater 7(3):1040–1049
Seif-Naraghi SB, Salvatore MA, Schup-Magoffin PJ et al (2010) Design and characterization of an injectable pericardial matrix gel: a potentially autologous scaffold for cardiac tissue engineering. Tissue Eng Part A 16(6):2017–2027
Freytes DO, Martin J, Velankar SS et al (2008) Preparation and rheological characterization of a gel form of the porcine urinary bladder matrix. Biomaterials 29(11):1630–1637
DeQuach JA, Yuan SH, Goldstein LS et al (2011) Decellularized porcine brain matrix for cell culture and tissue engineering scaffolds. Tissue Eng Part A 17(21–22):2583–2592
Wolf MT, Daly KA, Brennan-Pierce EP et al (2012) A hydrogel derived from decellularized dermal extracellular matrix. Biomaterials 33(29):7028–7038
Acknowledgments
A special thanks to Matt Wolf, Janet Reing, and Scott Johnson of the McGowan Institute for Regenerative Medicine for their assistance with technical details for the methods section.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Brown, B.N., Buckenmeyer, M.J., Prest, T.A. (2017). Preparation of Decellularized Biological Scaffolds for 3D Cell Culture. 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_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7021-6_2
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7019-3
Online ISBN: 978-1-4939-7021-6
eBook Packages: Springer Protocols