Dataset Information

Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes.

ABSTRACT: Stem cell-derived somatic cells represent an unlimited resource for basic and translational science. Although promising, there are significant hurdles that must be overcome. Our focus is on the generation of the major cell type of the human liver, the hepatocyte. Current protocols produce variable populations of hepatocytes that are the product of using undefined components in the differentiation process. This serves as a significant barrier to scale-up and application. To tackle this issue, we designed a defined differentiation process using recombinant laminin substrates to provide instruction. We demonstrate efficient hepatocyte specification, cell organization, and significant improvements in cell function and phenotype. This is driven in part by the suppression of unfavorable gene regulatory networks that control cell proliferation and migration, pluripotent stem cell self-renewal, and fibroblast and colon specification. We believe that this represents a significant advance, moving stem cell-based hepatocytes closer toward biomedical application.

SUBMITTER: Cameron K

PROVIDER: S-EPMC4682209 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes.

Cameron Kate K Tan Rosanne R Schmidt-Heck Wolfgang W Campos Gisela G Lyall Marcus J MJ Wang Yu Y Lucendo-Villarin Baltasar B Szkolnicka Dagmara D Bates Nicola N Kimber Susan J SJ Hengstler Jan G JG Godoy Patricio P Forbes Stuart J SJ Hay David C DC

Stem cell reports 20151125 6

Stem cell-derived somatic cells represent an unlimited resource for basic and translational science. Although promising, there are significant hurdles that must be overcome. Our focus is on the generation of the major cell type of the human liver, the hepatocyte. Current protocols produce variable populations of hepatocytes that are the product of using undefined components in the differentiation process. This serves as a significant barrier to scale-up and application. To tackle this issue, we ...[more]

PMID: 26626180

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Project description:Refined methods for maintaining specific functions of isolated hepatocytes under xeno-free and chemical defined conditions is of great importance for the development of hepatocyte research and regenerative therapy. Laminins, a large family of heterotrimeric basement membrane adhesion proteins, are highly cell and tissue type specific components of the extracellular matrix and strongly influence the behavior and function of associated cells and/or tissues. However, detailed biological functions of many laminin isoforms are still to be evaluated. In this study, we determined the distribution of laminin isoforms in human liver tissue and isolated primary human hepatocytes by western blot analysis, and investigated the efficacy of different human recombinant laminin isoforms on hepatic functions during culture. Protein expressions of laminin-chain ?2, ?3, ?4, ?1, ?3, ?1, and ?2 were detected in both isolated human hepatocytes and liver tissue. No ?1 and ?5 expression could be detected in liver tissue or hepatocytes. Hepatocytes were isolated from five different individual livers, and cultured on human recombinant laminin isoforms -111, -211, -221, -332, -411, -421, -511, and -521 (Biolamina AB), matrigel (extracted from Engelbreth-Holm-Swarm sarcoma), or collagen type IV (Collagen). Hepatocytes cultured on laminin showed characteristic hexagonal shape in a flat cell monolayer. Viability, double stranded DNA concentration, and Ki67 expression for hepatocytes cultured for six days on laminin were comparable to those cultured on EHS and Collagen. Hepatocytes cultured on laminin also displayed production of human albumin, alpha-1-antitrypsin, bile acids, and gene expression of liver-enriched factors, such as hepatocyte nuclear factor 4 alpha, glucose-6-phosphate, cytochrome P450 3A4, and multidrug resistance-associated protein 2. We conclude that all forms of human recombinant laminin tested maintain cell viability and liver-specific functions of primary human hepatocytes, and that recombinant laminin is a promising xeno-free and chemical defined strategy for preservation of hepatocyte specific function in vitro.

Dataset Information

Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes.

Publications

Recombinant Laminins Drive the Differentiation and Self-Organization of hESC-Derived Hepatocytes.

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