Functional Hepatocyte-Like Cells from Human Pluripotent Stem Cells in a Scalable Suspension Culture: A Success Story 2011-2018

Background and Aim: Biomedical application of human pluripotentstem cell-derived hepatocyte-like cells relies on efficient large-scaledifferentiation, which performed by a suspension culture of threedimensionalmulticellular spheroids in bioreactors.Methods: In this wok, we report on a scalable stirred-suspensionbioreactor culture of functional hepatocyte-like cells from the humanpluripotent stem cells. Three PhD projects designed to address thepotential challenges in this subject. This method needs large amounts ofgrowth factors and must overcome the limited diffusional transport posedby the inherent 3D structure of hPSC spheroids.Results: Currently used protocols for the scalable and integrateddifferentiation of hPSCs limited due to the high cost of growth factorsand technical challenges. Priming hPSCs aggregates with different smallmolecules in combination with different concentrations of growth factorsresulted in an effective protocol for differentiation. Hepatocyte-like cellscharacterized and then purified based on their physiological function.The differentiated HLCs possessed multiple characteristics of theprimary hepatocytes, including the expression patterns of liver-specificmarker genes, albumin secretion, urea production, collagen synthesis,indocyanine green and LDL uptake, glycogen storage, and induciblecytochrome P450 activity.Conclusion: These findings imply that sustained and localized deliveryof GFs using MPs might provide a de novo method towards scalabletechnologies for the hepatocytic differentiation and engineer a better 3Dmicroenvironment for cells.

کلیدواژه ها:

Hepatocyte-like cells ، Pluripotent stem cells ، Scalable suspension culture