Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding

Panagiotis Maghsoudlou; Fanourios Georgiades; Holly Smith; Anna Milan; Panicos Shangaris; Luca Urbani; Stavros P. Loukogeorgakis; Benedetta Lombardi; Giuseppe Mazza; Charlotte Hagen; Neil J. Sebire; Mark Turmaine; Simon Eaton; Alessandro Olivo; Jasminka Godovac-Zimmermann; Massimo Pinzani; Paul Gissen; Paolo De Coppi

doi:10.1371/journal.pone.0155324

Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding

Panagiotis Maghsoudlou, Fanourios Georgiades, Holly Smith, Anna Milan, Panicos Shangaris, Luca Urbani, Stavros P. Loukogeorgakis, Benedetta Lombardi, Giuseppe Mazza, Charlotte Hagen, Neil J. Sebire, Mark Turmaine, Simon Eaton, Alessandro Olivo, Jasminka Godovac-Zimmermann, Massimo Pinzani, Paul Gissen, Paolo De Coppi

Women & Children's Health

Research output: Contribution to journal › Article › peer-review

74 Citations (Scopus)

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.

Original language	English
Article number	e0155324
Journal	PLoS ONE
Volume	11
Issue number	5
DOIs	https://doi.org/10.1371/journal.pone.0155324
Publication status	Published - 9 May 2016

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Maghsoudlou, P., Georgiades, F., Smith, H., Milan, A., Shangaris, P., Urbani, L., Loukogeorgakis, S. P., Lombardi, B., Mazza, G., Hagen, C., Sebire, N. J., Turmaine, M., Eaton, S., Olivo, A., Godovac-Zimmermann, J., Pinzani, M., Gissen, P., & De Coppi, P. (2016). Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding. PLoS ONE, 11(5), Article e0155324. https://doi.org/10.1371/journal.pone.0155324

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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.",

author = "Panagiotis Maghsoudlou and Fanourios Georgiades and Holly Smith and Anna Milan and Panicos Shangaris and Luca Urbani and Loukogeorgakis, {Stavros P.} and Benedetta Lombardi and Giuseppe Mazza and Charlotte Hagen and Sebire, {Neil J.} and Mark Turmaine and Simon Eaton and Alessandro Olivo and Jasminka Godovac-Zimmermann and Massimo Pinzani and Paul Gissen and {De Coppi}, Paolo",

year = "2016",

month = may,

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doi = "10.1371/journal.pone.0155324",

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Maghsoudlou, P, Georgiades, F, Smith, H, Milan, A, Shangaris, P, Urbani, L, Loukogeorgakis, SP, Lombardi, B, Mazza, G, Hagen, C, Sebire, NJ, Turmaine, M, Eaton, S, Olivo, A, Godovac-Zimmermann, J, Pinzani, M, Gissen, P & De Coppi, P 2016, 'Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding', PLoS ONE, vol. 11, no. 5, e0155324. https://doi.org/10.1371/journal.pone.0155324

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T1 - Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding

AU - Maghsoudlou, Panagiotis

AU - Georgiades, Fanourios

AU - Smith, Holly

AU - Milan, Anna

AU - Shangaris, Panicos

AU - Urbani, Luca

AU - Loukogeorgakis, Stavros P.

AU - Lombardi, Benedetta

AU - Mazza, Giuseppe

AU - Hagen, Charlotte

AU - Sebire, Neil J.

AU - Turmaine, Mark

AU - Eaton, Simon

AU - Olivo, Alessandro

AU - Godovac-Zimmermann, Jasminka

AU - Pinzani, Massimo

AU - Gissen, Paul

AU - De Coppi, Paolo

PY - 2016/5/9

Y1 - 2016/5/9

N2 - 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.

AB - 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.

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U2 - 10.1371/journal.pone.0155324

DO - 10.1371/journal.pone.0155324

M3 - Article

C2 - 27159223

AN - SCOPUS:84969790475

SN - 1932-6203

VL - 11

JO - PLoS ONE

JF - PLoS ONE

IS - 5

M1 - e0155324

ER -

Optimization of liver decellularization maintains extracellular matrix micro-architecture and composition predisposing to effective cell seeding

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