title: ECM proteins in a microporous scaffold influence hepatocyte morphology, function, and gene expression.
abstract: It is well known that a three-dimensional (3D) culture environment and the presence of extracellular matrix (ECM) proteins facilitate hepatocyte viability and maintenance of the liver-specific phenotype in vitro. However, it is not clear whether specific ECM components such as collagen or fibronectin differentially regulate such processes, especially in 3D scaffolds. In this study, a series of ECM-functionalized inverted colloidal crystal (ICC) microporous scaffolds were fabricated and their influence on Huh-7.5 cell proliferation, morphology, hepatic-specific functions, and patterns of gene expression were compared. Both collagen and fibronectin promoted albumin production and liver-specific gene expression of Huh-7.5 cells, compared with the bare ICC scaffold. Interestingly, cells in the fibronectin-functionalized scaffold exhibited different aggregation patterns to those in the collagen-functionalized scaffold, a variation that could be related to the distinct mRNA expression levels of cell adhesion-related genes. Based on these results, we can conclude that different ECM proteins, such as fibronectin and collagen, indeed play distinct roles in the phenotypic regulation of cells cultured in a 3D environment.
authors.affiliation: School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore.|Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive, 637553, Singapore.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore.|Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive, 637553, Singapore.
authors.forename: Myung Hee
authors.initials: MH
authors.lastname: Kim
authors.affiliation: School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore.|Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive, 637553, Singapore.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Shirahama
authors.affiliation: School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore.|Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive, 637553, Singapore.
authors.forename: Jae Ho
authors.initials: JH
authors.lastname: Lee
authors.affiliation: School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore.|Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive, 637553, Singapore.
authors.forename: Soon Seng
authors.initials: SS
authors.lastname: Ng
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Medicine, Stanford University School of Medicine, Palo Alto, California, United States of America.|Department of Microbiology and Immunology, Stanford University School of Medicine, Palo Alto, California, United States of America.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Glenn
authors.affiliation: School of Materials Science and Engineering, Nanyang Technological University, 50 Nanyang Avenue, 639798, Singapore.|Centre for Biomimetic Sensor Science, Nanyang Technological University, 50 Nanyang Drive, 637553, Singapore.|School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Avenue, 637459, Singapore.
authors.forename: Nam-Joon
authors.initials: NJ
authors.lastname: Cho
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000418:Albumins / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D024042:Collagen Type I / Q000737:chemistry / Q000494:pharmacology*; D003102:Colloids; D051544:Cytochrome P-450 CYP3A / Q000235:genetics / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry; D005353:Fibronectins / Q000737:chemistry / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D051557:Hepatocyte Nuclear Factor 4 / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D016023:Integrins / Q000235:genetics / Q000378:metabolism; D062793:Occludin / Q000235:genetics / Q000378:metabolism; D016062:Porosity; D054457:Tissue Scaffolds
chemical_list: D000199:Actins; D000418:Albumins; D015820:Cadherins; D024042:Collagen Type I; D003102:Colloids; D005353:Fibronectins; C496592:HNF4A protein, human; D051557:Hepatocyte Nuclear Factor 4; D016023:Integrins; C568985:OCLN protein, human; D062793:Occludin; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human