Enhanced growth and hepatic differentiation of fetal liver epithelial cells through combinational and temporal adjustment of soluble factors

Abstract

Fetal liver epithelial cells (FLEC) are valuable for liver cell therapy and tissue engineering, but methods for culture and characterization of these cells are not well developed. This work explores the influence of multiple soluble factors on FLEC, with the long-term goal of developing an optimal culture system to generate functional liver tissue. Our comparative analysis suggests hepatocyte growth factor (HGF) is required throughout the culture period. In the presence of HGF, addition of oncostatin M (OSM) at culture initiation results in concurrent growth and maturation, while constant presence of protective agents like ascorbic acid enhances cell survival. Study observations led to the development of a culture medium that provided optimal growth and hepatic differentiation conditions. FLEC expansion was observed to be ~2 fold of that under standard conditions, albumin secretion rate was 2 – 3 times greater than maximal values obtained with other media, and the highest level of glycogen accumulation among all conditions was observed with the developed medium. Our findings serve to advance culture methods for liver progenitors in cell therapy and tissue engineering applications.

High Efficient Differentiation of Functional Hepatocytes from Porcine Induced Pluripotent Stem Cells

“…BMPs, in parallel to FGF, have been reported as critical growth factors for the growth of hepatic endoderm [21] . Additionally, both HGF and EGF were found to be essential for hepatoblast proliferation [22] . Therefore, adding of BMP4, bFGF, HGF and EGF from differentiation day 6 to day 9 could substantially promote the expansion of early hepatic progenitor cells derived from piPSC-derived DE.…”

Unbalanced distribution of materials: the art of giving rise to hepatocytes from liver stem/progenitor cells

“…Because the maturation of LSPCs into the hepatocyte lineage is regulated by numerous factors, combining the above GFs can synergistically induce the hepatic differentiation of LSPCs. For example, the combination of HGF and OSM can more efficiently induce the hepatic differentiation of LSPCs 103 – 104 . Recently, some new stimulators have been identified as initiating the hepatic differentiation of LSPCs, such as the polycomb group protein Ezh2 105 and S-Adenosylmethionine (abbreviated as SAM, SAMe or AdoMet) 106 .…”

Extramedullary Hematopoiesis of the Liver and Spleen

“…In early hepatic hematopoiesis, the first cells that are generated are the undifferentiated mononuclear cells (future hematopoietic stem cells). The primitive erythroid cells are the most abundant cell lines found at this stage [6], but the presence of mast cells, and natural killer and innate lymphoid cell precursors in the yolk sac, were also detected [5]. The erythroblasts or megaloblasts produced at this stage are not pluripotent or self-renewing, because the main interest of the body, at this point, is to ensure rapid embryonic growth and development.…”

“…The yolk sac has a more immature, less differentiated type of erythropoiesis, where the starting cells are derived from the mesodermal cells, whereas in the liver, the cells of origin are hematopoietic stem cells [5]. Besides these, the liver is colonized with HSCs from a hemogenic endothelium on the ventral wall of the aorta [6]. In the liver, the cells of origin have a lesser difference of maturation between the nucleus and cytoplasm, and a smaller number of polyribosomes [3].…”

“…In the liver, the cells of origin have a lesser difference of maturation between the nucleus and cytoplasm, and a smaller number of polyribosomes [3]. The hepatic erythrocytes rapidly differentiate and take the place of the circulating HSCs found in early embryonic development [6]. In general, hepatic erythropoiesis has many similarities to bone marrow erythropoiesis.…”