Differences in definitive endoderm induction approaches using growth factors and small molecules

Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. Developing human liver cell models for pharmaceutical applications is highly demanding. Due to the vast number of existing protocols to generate DE cells from hPSCs, we aimed...

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Veröffentlicht in:Journal of cellular physiology 2018-04, Vol.233 (4), p.3578-3589
Hauptverfasser: Bogacheva, Mariia S., Khan, Sofia, Kanninen, Liisa K., Yliperttula, Marjo, Leung, Alan W., Lou, Yan‐Ru
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container_end_page 3589
container_issue 4
container_start_page 3578
container_title Journal of cellular physiology
container_volume 233
creator Bogacheva, Mariia S.
Khan, Sofia
Kanninen, Liisa K.
Yliperttula, Marjo
Leung, Alan W.
Lou, Yan‐Ru
description Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. Developing human liver cell models for pharmaceutical applications is highly demanding. Due to the vast number of existing protocols to generate DE cells from hPSCs, we aimed to compare the specificity and efficiency of selected published differentiation conditions. We differentiated two hPSC lines (induced PSC and embryonic stem cell) to DE cells on Matrigel matrix using growth factors (Activin A and Wnt‐3a) and small molecules (sodium butyrate and IDE 1) in different combinations. By studying dynamic changes during 6 days in cell morphology and the expression of markers for pluripotency, DE, and other germ layer lineages, we found that Activin A is essential for DE differentiation, while Wnt‐3a and sodium butyrate are dispensable. Although sodium butyrate exerted rapid DE differentiation kinetics, it caused massive cell death and could not generate sufficient cells for further differentiation and applications. We further discover that IDE 1 could not induce DE as reported previously. Hereby, we compared different conditions for DE induction and found an effective six day‐protocol to obtain DE cells for the further differentiation and applications. Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. We differentiated two hPSC lines using growth factors and small molecules in different combinations. We identified an efficient and robust method for generation of DE cells from hPSCs.
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Developing human liver cell models for pharmaceutical applications is highly demanding. Due to the vast number of existing protocols to generate DE cells from hPSCs, we aimed to compare the specificity and efficiency of selected published differentiation conditions. We differentiated two hPSC lines (induced PSC and embryonic stem cell) to DE cells on Matrigel matrix using growth factors (Activin A and Wnt‐3a) and small molecules (sodium butyrate and IDE 1) in different combinations. By studying dynamic changes during 6 days in cell morphology and the expression of markers for pluripotency, DE, and other germ layer lineages, we found that Activin A is essential for DE differentiation, while Wnt‐3a and sodium butyrate are dispensable. Although sodium butyrate exerted rapid DE differentiation kinetics, it caused massive cell death and could not generate sufficient cells for further differentiation and applications. We further discover that IDE 1 could not induce DE as reported previously. Hereby, we compared different conditions for DE induction and found an effective six day‐protocol to obtain DE cells for the further differentiation and applications. Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. We differentiated two hPSC lines using growth factors and small molecules in different combinations. 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We further discover that IDE 1 could not induce DE as reported previously. Hereby, we compared different conditions for DE induction and found an effective six day‐protocol to obtain DE cells for the further differentiation and applications. Definitive endoderm (DE) is the first stage of human pluripotent stem cell (hPSC) differentiation into hepatocyte‐like cells. We differentiated two hPSC lines using growth factors and small molecules in different combinations. 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subjects Activin
Activins - pharmacology
Butyric Acid - pharmacology
Cell culture
Cell death
cell differentiation
Cell Differentiation - drug effects
Cell Line
Cell morphology
Cells, Cultured
Cytology
definitive endoderm
Differentiation (biology)
Embryonic Stem Cells - cytology
Embryonic Stem Cells - drug effects
Endoderm
Endoderm - cytology
Endoderm - drug effects
Growth factors
Hepatocytes
Hepatocytes - drug effects
Hepatocytes - metabolism
human pluripotent stem cell
Humans
Liver
Pluripotency
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - drug effects
Sodium
Sodium butyrate
Stem cells
Wnt protein
title Differences in definitive endoderm induction approaches using growth factors and small molecules
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