Abstract
Lately, the need for three-dimensional (3D) cell culture has been recognized in order to closely mimic the organization of native tissues. Thus, 3D scaffolds started to be employed to facilitate the 3D cell organization and enable the artificial tissue formation for the emerging tissue engineering applications. 3D scaffolds can be prepared by various techniques, each with certain advantages and disadvantages. Decellularization is an easy method based on removal of cells from native tissue sample, yielding extracellular matrix (ECM) scaffold with preserved architecture and bioactivity. This chapter provides a detailed protocol for decellularization of pig lung and also some basic assays for evaluation of its effectivity, such as determination of DNA content and histological verification of the selected ECM components. Such decellularized scaffold can subsequently be used for various tissue engineering applications, for example, for recellularization with cells of interest, for natural ECM hydrogel preparation, or as a bioink for 3D bioprinting.
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References
Langer R, Vacanti JP (1993) Tissue engineering. Science 260:920–926
Sedláková V, Kloučková M, Garlíková Z et al (2019) Options for modeling the respiratory system: inserts, scaffolds and microfluidic chips. Drug Discov Today 24:971–982
Laurencin CT, Nair LS (eds) (2008) Nanotechnology and tissue engineering: the scaffold. CRC Press, Boca Raton
Sedlakova V, Ruel M, Suuronen EJ (2019) Therapeutic use of bioengineered materials for myocardial infarction. In: Alarcon EI, Ahumada M (eds) Nanoengineering materials for biomedical uses. Springer, Cham, pp 161–193
Boland ED, Espy PG, Bowlin GL (2008) Tissue engineering scaffolds. In: Wnek GE, Bowlin GL (eds) Encyclopedia of biomaterials and biomedical engineering. Informa Healthcare USA, Inc, New York, pp 2828–2837
Hasirci V, Yucel D (2008) Polymers used in tissue engineering. In: Wnek GE, Bowlin GL (eds) Encyclopedia of biomaterials and biomedical engineering. Informa Healthcare USA, Inc, New York, pp 2282–2299
Rajab TK, O’malley TJ, Tchantchaleishvili V (2020) Decellularized scaffolds for tissue engineering: current status and future perspective. Artif Organs 44:1031–1043
Price AP, Godin LM, Domek A et al (2015) Automated decellularization of intact, human-sized lungs for tissue engineering. Tissue Eng Part C Methods 21:94–103
Garlíková Z, Silva AC, Rabata A et al (2018) Generation of a close-to-native in vitro system to study lung cells-extracellular matrix crosstalk. Tissue Eng Part C Methods 24:1–13
Song JJ, Kim SS, Liu Z et al (2011) Enhanced in vivo function of bioartificial lungs in rats. Ann Thorac Surg 92:998–1006
Ott HC, Clippinger B, Conrad C et al (2010) Regeneration and orthotopic transplantation of a bioartificial lung. Nat Med 16:927–933
Henwood A (2002) The orcein stain – a versatile stain for histopathology. J Histotechnol 25:29–31
Van Kuppevelt TH, Veerkamp JH, Timmermans JA (1995) Immunoquantification of type I, III, IV and V collagen in small samples of human lung parenchyma. Int J Biochem Cell Biol 27:775–782
De Hilster RHJ, Sharma PK, Jonker MR et al (2020) Human lung extracellular matrix hydrogels resemble the stiffness and viscoelasticity of native lung tissue. Am J Phys Lung Cell Mol Phys 318:L698–l704
Pouliot RA, Young BM, Link PA et al (2020) Porcine lung-derived extracellular matrix hydrogel properties are dependent on pepsin digestion time. Tissue Eng Part C Methods 26:332–346
Kim BS, Kim H, Gao G et al (2017) Decellularized extracellular matrix: a step towards the next generation source for bioink manufacturing. Biofabrication 9:034104
Acknowledgments
This work was supported by the Czech Science Foundation (grant no. 18-00145S), the Ministry of Health of the Czech Republic (grant no. 16-31501A), Masaryk University (MUNI/A/1398/2021), and the European Regional Development Fund – Project INBIO (No. CZ.02.1.01/0.0/0.0/16_026/0008451).
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Čimborová, K., Kotasová, H., Pelková, V., Sedláková, V., Hampl, A. (2024). Decellularization of Pig Lung to Yield Three-Dimensional Scaffold for Lung Tissue Engineering. In: Sumbalova Koledova, Z. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 2764. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3674-9_3
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DOI: https://doi.org/10.1007/978-1-0716-3674-9_3
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