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Article

A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture

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Department of Medicine, Division of Respirology, Firestone Institute for Respiratory Health, McMaster University, Hamilton, ON L8N 4A6, Canada
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Department of Chemistry and Chemical Biology, McMaster University, Hamilton, ON L8S 4M1, Canada
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Department of Chemical Engineering, McMaster University, Hamilton, ON L8S 4L7, Canada
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School of Biomedical Engineering, McMaster University, Hamilton, ON L8S 4K1, Canada
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McMaster Immunology Research Centre, Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON L8S 4K1, Canada
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Department of Surgery, McMaster University, Hamilton, ON L8S 4K1, Canada
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Division of Respiratory Medicine, Department of Medicine, University of British Columbia, Vancouver, BC V6H 3Z6, Canada
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Department of Biology, University of Waterloo, Waterloo, ON N2L 3G1, Canada
*
Author to whom correspondence should be addressed.
Academic Editors: Tillie-Louise Hackett and Emmanuel Twumasi Osei
Cells 2021, 10(6), 1538; https://doi.org/10.3390/cells10061538
Received: 31 May 2021 / Revised: 11 June 2021 / Accepted: 15 June 2021 / Published: 18 June 2021
Decellularization efforts must balance the preservation of the extracellular matrix (ECM) components while eliminating the nucleic acid and cellular components. Following effective removal of nucleic acid and cell components, decellularized ECM (dECM) can be solubilized in an acidic environment with the assistance of various enzymes to develop biological scaffolds in different forms, such as sheets, tubular constructs, or three-dimensional (3D) hydrogels. Each organ or tissue that undergoes decellularization requires a distinct and optimized protocol to ensure that nucleic acids are removed, and the ECM components are preserved. The objective of this study was to optimize the decellularization process for dECM isolation from human lung tissues for downstream 2D and 3D cell culture systems. Following protocol optimization and dECM isolation, we performed experiments with a wide range of dECM concentrations to form human lung dECM hydrogels that were physically stable and biologically responsive. The dECM based-hydrogels supported the growth and proliferation of primary human lung fibroblast cells in 3D cultures. The dECM is also amenable to the coating of polyester membranes in Transwell™ Inserts to improve the cell adhesion, proliferation, and barrier function of primary human bronchial epithelial cells in 2D. In conclusion, we present a robust protocol for human lung decellularization, generation of dECM substrate material, and creation of hydrogels that support primary lung cell viability in 2D and 3D culture systems View Full-Text
Keywords: decellularization; lung; cell culture; hydrogels; coating; epithelial; fibroblast decellularization; lung; cell culture; hydrogels; coating; epithelial; fibroblast
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MDPI and ACS Style

Dabaghi, M.; Saraei, N.; Carpio, M.B.; Nanduri, V.; Ungureanu, J.; Babi, M.; Chandiramohan, A.; Noble, A.; Revill, S.D.; Zhang, B.; Ask, K.; Kolb, M.; Shargall, Y.; Moran-Mirabal, J.; Hirota, J.A. A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture. Cells 2021, 10, 1538. https://doi.org/10.3390/cells10061538

AMA Style

Dabaghi M, Saraei N, Carpio MB, Nanduri V, Ungureanu J, Babi M, Chandiramohan A, Noble A, Revill SD, Zhang B, Ask K, Kolb M, Shargall Y, Moran-Mirabal J, Hirota JA. A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture. Cells. 2021; 10(6):1538. https://doi.org/10.3390/cells10061538

Chicago/Turabian Style

Dabaghi, Mohammadhossein, Neda Saraei, Mabel Barreiro Carpio, Vibudha Nanduri, Julia Ungureanu, Mouhanad Babi, Abiram Chandiramohan, Alexander Noble, Spencer D. Revill, Boyang Zhang, Kjetil Ask, Martin Kolb, Yaron Shargall, Jose Moran-Mirabal, and Jeremy Alexander Hirota. 2021. "A Robust Protocol for Decellularized Human Lung Bioink Generation Amenable to 2D and 3D Lung Cell Culture" Cells 10, no. 6: 1538. https://doi.org/10.3390/cells10061538

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