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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-05-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/6/1357 |
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Article |
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DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Niki Alevra Sarika Valéry L. Payen Maximilien Fléron Joachim Ravau Davide Brusa Mustapha Najimi Edwin De Pauw Gauthier Eppe Gabriel Mazzucchelli Etienne M. Sokal Anne des Rieux Adil El Taghdouini |
| spellingShingle |
Niki Alevra Sarika Valéry L. Payen Maximilien Fléron Joachim Ravau Davide Brusa Mustapha Najimi Edwin De Pauw Gauthier Eppe Gabriel Mazzucchelli Etienne M. Sokal Anne des Rieux Adil El Taghdouini Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver Cells Cells liver primary cells extracellular matrix decellularization proteomics |
| author_facet |
Niki Alevra Sarika Valéry L. Payen Maximilien Fléron Joachim Ravau Davide Brusa Mustapha Najimi Edwin De Pauw Gauthier Eppe Gabriel Mazzucchelli Etienne M. Sokal Anne des Rieux Adil El Taghdouini |
| author_sort |
Niki Alevra Sarika |
| title |
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_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_sort |
human liver-derived extracellular matrix for the culture of distinct human primary liver cells |
| publisher |
MDPI AG |
| series |
Cells |
| issn |
2073-4409 |
| publishDate |
2020-05-01 |
| 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. |
| topic |
liver primary cells extracellular matrix decellularization proteomics |
| url |
https://www.mdpi.com/2073-4409/9/6/1357 |
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doaj-f090be8d1fc34fab9b13f13685419d5c2020-11-25T03:17:33ZengMDPI AGCells2073-44092020-05-0191357135710.3390/cells9061357Human Liver-Derived Extracellular Matrix for the Culture of Distinct Human Primary Liver CellsNiki Alevra Sarika0Valéry L. Payen1Maximilien Fléron2Joachim Ravau3Davide Brusa4Mustapha Najimi5Edwin De Pauw6Gauthier Eppe7Gabriel Mazzucchelli8Etienne M. Sokal9Anne des Rieux10Adil El Taghdouini11Laboratory of Pediatric Hepatology and Cell Therapy (PEDI), Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Mounier 52 Box B1.52.03, 1200 Brussels, BelgiumLaboratory of Pediatric Hepatology and Cell Therapy (PEDI), Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Mounier 52 Box B1.52.03, 1200 Brussels, BelgiumGIGA-Proteomics Facility, University of Liège (ULiège), Allée du six août 11, 4000 Liège, BelgiumLaboratory of Pediatric Hepatology and Cell Therapy (PEDI), Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Mounier 52 Box B1.52.03, 1200 Brussels, BelgiumFlow Cytometry Technological Platform, Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Hippocrate 55 Box B1.55.20, 1200 Brussels, BelgiumLaboratory of Pediatric Hepatology and Cell Therapy (PEDI), Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Mounier 52 Box B1.52.03, 1200 Brussels, BelgiumMass Spectrometry Laboratory (MSLab), MolSys Research Unit, University of Liège (ULiège), Allée du six août 11, 4000 Liège, BelgiumMass Spectrometry Laboratory (MSLab), MolSys Research Unit, University of Liège (ULiège), Allée du six août 11, 4000 Liège, BelgiumMass Spectrometry Laboratory (MSLab), MolSys Research Unit, University of Liège (ULiège), Allée du six août 11, 4000 Liège, BelgiumLaboratory of Pediatric Hepatology and Cell Therapy (PEDI), Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Mounier 52 Box B1.52.03, 1200 Brussels, BelgiumLaboratory of Advanced Drug Delivery and Biomaterials (ADDB), Louvain Drug Research Institute (LDRI), Université catholique de Louvain (UCLouvain), Avenue Mounier 73 Box B1.73.12, 1200 Brussels, BelgiumLaboratory of Pediatric Hepatology and Cell Therapy (PEDI), Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain (UCLouvain), Avenue Mounier 52 Box B1.52.03, 1200 Brussels, BelgiumThe 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.https://www.mdpi.com/2073-4409/9/6/1357liverprimary cellsextracellular matrixdecellularizationproteomics |