Co-culture with mouse embryonic fibroblasts improves maintenance of metabolic function of human small hepatocyte progenitor cells

Abstract

Derivation and culture of small hepatocyte progenitor cells (SHPCs) capable of proliferating in vitro has been described in rodents and recently in humans. These cells are capable of engrafting in injured livers, however, they display de-differentiated morphology and reduced xenobiotic metabolism activity in culture over passages. Here we report that SHPCs derived from adult primary human hepatocytes (PHHs) and cultured on mouse embryonic fibroblasts (MEFs) not only display differentiated morphology and exhibit gene expression profiles similar to adult PHHs, but importantly, they retain their phenotype over several passages. Further, unlike previous reports, where extensive manipulations of culture conditions are required to convert SHPCs to metabolically functional hepatocytes, SHPCs in our co-culture system maintain expression of xenobiotic metabolism-associated genes. We show that SHPCs in co-culture are able to perform xenobiotic metabolism at rates equal to their parent PHHs as evidenced by the metabolism of acetaminophen to all of its major metabolites. In summary, we present an improved co-culture system that allows generation of SHPCs from adult PHHs that maintain their differentiated phenotype over multiple passages. Our findings would be useful for expansion of limited PHHs for use in studies of drug metabolism and toxicity testing.

Growing Role of 3D In Vitro Cell Cultures in the Study of Cellular and Molecular Mechanisms: Short Focus on Breast Cancer, Endometriosis, Liver and Infectious Diseases

“…Additionally, the strategies currently adopted for fabricating 3D cultures enabled the production of multicellular cocultures. These tools allowed for exploring the impact providing cultured cells with relevant cell–cell interactions and underlined a further boost of the metabolic activity of hepatocyte-like cells when co-cultured with other non-parenchymal cell types (e.g., fibroblasts, stellate cells, endothelial cells) [ 107 , 108 , 109 , 110 , 111 ]. 3D cell cultures emerged as a powerful tool for enhancing the phenotypic stability of both primary hepatocytes and iPSC-derived hepatocytes ( Figure 3 c).…”

CINC-2 and miR-199a-5p in EVs secreted by transplanted Thy1+ cells activate hepatocytic progenitor cell growth in rat liver regeneration

“…Small hepatocytes (SHs) are a subpopulation of hepatocytes found in rats, mice, and humans. SHs isolated from healthy adult rat [ 12 , 13 ], mouse [ 14 ], and human [ 15 , 16 ] livers can clonally proliferate to form colonies and differentiate into mature hepatocytes (MHs) in vitro. We previously identified CD44 as a marker of SHs [ 17 ].…”