title: Optimization of Liver Decellularization Maintains Extracellular Matrix Micro-Architecture and Composition Predisposing to Effective Cell Seeding.
abstract: Hepatic tissue engineering using decellularized scaffolds is a potential therapeutic alternative to conventional transplantation. However, scaffolds are usually obtained using decellularization protocols that destroy the extracellular matrix (ECM) and hamper clinical translation. We aim to develop a decellularization technique that reliably maintains hepatic microarchitecture and ECM components. Isolated rat livers were decellularized by detergent-enzymatic technique with (EDTA-DET) or without EDTA (DET). Histology, DNA quantification and proteomics confirmed decellularization with further DNA reduction with the addition of EDTA. Quantification, histology, immunostaining, and proteomics demonstrated preservation of extracellular matrix components in both scaffolds with a higher amount of collagen and glycosaminoglycans in the EDTA-DET scaffold. Scanning electron microscopy and X-ray phase contrast imaging showed microarchitecture preservation, with EDTA-DET scaffolds more tightly packed. DET scaffold seeding with a hepatocellular cell line demonstrated complete repopulation in 14 days, with cells proliferating at that time. Decellularization using DET preserves microarchitecture and extracellular matrix components whilst allowing for cell growth for up to 14 days. Addition of EDTA creates a denser, more compact matrix. Transplantation of the scaffolds and scaling up of the methodology are the next steps for successful hepatic tissue engineering.
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Maghsoudlou
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Fanourios
authors.initials: F
authors.lastname: Georgiades
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London, WC1E 6BT, United Kingdom.
authors.forename: Holly
authors.initials: H
authors.lastname: Smith
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Milan
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Panicos
authors.initials: P
authors.lastname: Shangaris
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Luca
authors.initials: L
authors.lastname: Urbani
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Stavros P
authors.initials: SP
authors.lastname: Loukogeorgakis
authors.affiliation: London Center for Nephrology, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Lombardi
authors.affiliation: Institute for Liver and Digestive Health, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Mazza
authors.affiliation: Department of Medical Physics & Bioengineering, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Hagen
authors.affiliation: Department of Histopathology, UCL Institute of Child Health and Great Ormond Street Hospital, London, WC1N 1EH, United Kingdom.
authors.forename: Neil J
authors.initials: NJ
authors.lastname: Sebire
authors.affiliation: Division of Bioscience, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Mark
authors.initials: M
authors.lastname: Turmaine
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Simon
authors.initials: S
authors.lastname: Eaton
authors.affiliation: Department of Medical Physics & Bioengineering, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Olivo
authors.affiliation: London Center for Nephrology, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Jasminka
authors.initials: J
authors.lastname: Godovac-Zimmermann
authors.affiliation: Institute for Liver and Digestive Health, University College London, London, WC1N 1EH, United Kingdom.
authors.forename: Massimo
authors.initials: M
authors.lastname: Pinzani
authors.affiliation: MRC Laboratory for Molecular Cell Biology, University College London, London, WC1E 6BT, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Gissen
authors.affiliation: Department of Stem Cells and Regenerative Medicine, UCL Institute of Child Health and Great Ormond Street Hospital, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
authors.forename: Paolo
authors.initials: P
authors.lastname: De Coppi
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D056945:Hep G2 Cells; D006801:Humans; D008099:Liver; D008855:Microscopy, Electron, Scanning; D051381:Rats; D053719:Tandem Mass Spectrometry; D054457:Tissue Scaffolds