Cargando…

Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells

The lack of robust methods to preserve, purify and in vitro maintain the phenotype of the human liver’s highly specialized parenchymal and non-parenchymal cell types importantly hampers their exploitation for the development of research and clinical applications. There is in this regard a growing in...

Descripción completa

Detalles Bibliográficos
Autores principales: Alevra Sarika, Niki, Payen, Valéry L., Fléron, Maximilien, Ravau, Joachim, Brusa, Davide, Najimi, Mustapha, De Pauw, Edwin, Eppe, Gauthier, Mazzucchelli, Gabriel, Sokal, Etienne M., des Rieux, Anne, El Taghdouini, Adil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7349413/
https://www.ncbi.nlm.nih.gov/pubmed/32486126
http://dx.doi.org/10.3390/cells9061357
_version_ 1783557058333769728
author Alevra Sarika, Niki
Payen, Valéry L.
Fléron, Maximilien
Ravau, Joachim
Brusa, Davide
Najimi, Mustapha
De Pauw, Edwin
Eppe, Gauthier
Mazzucchelli, Gabriel
Sokal, Etienne M.
des Rieux, Anne
El Taghdouini, Adil
author_facet Alevra Sarika, Niki
Payen, Valéry L.
Fléron, Maximilien
Ravau, Joachim
Brusa, Davide
Najimi, Mustapha
De Pauw, Edwin
Eppe, Gauthier
Mazzucchelli, Gabriel
Sokal, Etienne M.
des Rieux, Anne
El Taghdouini, Adil
author_sort Alevra Sarika, Niki
collection PubMed
description The lack of robust methods to preserve, purify and in vitro maintain the phenotype of the human liver’s highly specialized parenchymal and non-parenchymal cell types importantly hampers their exploitation for the development of research and clinical applications. There is in this regard a growing interest in the use of tissue-specific extracellular matrix (ECM) to provide cells with an in vitro environment that more closely resembles that of the native tissue. In the present study, we have developed a method that allows for the isolation and downstream application of the human liver’s main cell types from cryopreserved material. We also isolated and solubilized human liver ECM (HL-ECM), analyzed its peptidomic and proteomic composition by mass spectrometry and evaluated its interest for the culture of distinct primary human liver cells. Our analysis of the HL-ECM revealed proteomic diversity, type 1 collagen abundance and partial loss of integrity following solubilization. Solubilized HL-ECM was evaluated either as a coating or as a medium supplement for the culture of human primary hepatocytes, hepatic stellate cells and liver sinusoidal endothelial cells. Whereas the solubilized HL-ECM was suitable for cell culture, its impact on the phenotype and/or functionality of the human liver cells was limited. Our study provides a first detailed characterization of solubilized HL-ECM and a first report of its influence on the culture of distinct human primary liver cells.
format Online
Article
Text
id pubmed-7349413
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-73494132020-07-14 Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells Alevra Sarika, Niki Payen, Valéry L. Fléron, Maximilien Ravau, Joachim Brusa, Davide Najimi, Mustapha De Pauw, Edwin Eppe, Gauthier Mazzucchelli, Gabriel Sokal, Etienne M. des Rieux, Anne El Taghdouini, Adil Cells Article The lack of robust methods to preserve, purify and in vitro maintain the phenotype of the human liver’s highly specialized parenchymal and non-parenchymal cell types importantly hampers their exploitation for the development of research and clinical applications. There is in this regard a growing interest in the use of tissue-specific extracellular matrix (ECM) to provide cells with an in vitro environment that more closely resembles that of the native tissue. In the present study, we have developed a method that allows for the isolation and downstream application of the human liver’s main cell types from cryopreserved material. We also isolated and solubilized human liver ECM (HL-ECM), analyzed its peptidomic and proteomic composition by mass spectrometry and evaluated its interest for the culture of distinct primary human liver cells. Our analysis of the HL-ECM revealed proteomic diversity, type 1 collagen abundance and partial loss of integrity following solubilization. Solubilized HL-ECM was evaluated either as a coating or as a medium supplement for the culture of human primary hepatocytes, hepatic stellate cells and liver sinusoidal endothelial cells. Whereas the solubilized HL-ECM was suitable for cell culture, its impact on the phenotype and/or functionality of the human liver cells was limited. Our study provides a first detailed characterization of solubilized HL-ECM and a first report of its influence on the culture of distinct human primary liver cells. MDPI 2020-05-30 /pmc/articles/PMC7349413/ /pubmed/32486126 http://dx.doi.org/10.3390/cells9061357 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Alevra Sarika, Niki
Payen, Valéry L.
Fléron, Maximilien
Ravau, Joachim
Brusa, Davide
Najimi, Mustapha
De Pauw, Edwin
Eppe, Gauthier
Mazzucchelli, Gabriel
Sokal, Etienne M.
des Rieux, Anne
El Taghdouini, Adil
Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
title Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
title_full Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
title_fullStr Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
title_full_unstemmed Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
title_short Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells
title_sort human liver-derived extracellular matrix for the culture of distinct human primary liver cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7349413/
https://www.ncbi.nlm.nih.gov/pubmed/32486126
http://dx.doi.org/10.3390/cells9061357
work_keys_str_mv AT alevrasarikaniki humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT payenvaleryl humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT fleronmaximilien humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT ravaujoachim humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT brusadavide humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT najimimustapha humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT depauwedwin humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT eppegauthier humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT mazzucchelligabriel humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT sokaletiennem humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT desrieuxanne humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells
AT eltaghdouiniadil humanliverderivedextracellularmatrixforthecultureofdistincthumanprimarylivercells