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Generation of functional cholangiocyte-like cells from human pluripotent stem cells and HepaRG cells.

Dianat N, Dubois-Pot-Schneider H, Steichen C, Desterke C, Leclerc P, Raveux A, Combettes L, Weber A, Corlu A, Dubart-Kupperschmitt A - Hepatology (2014)

Bottom Line: In addition, we showed that cholangiocyte-like cells could also be generated from human induced pluripotent stem cells, demonstrating the efficacy of our approach with stem/progenitor cells of diverse origins.We have developed a robust and efficient method for differentiating pluripotent stem cells into cholangiocyte-like cells, which display structural and functional similarities to bile duct cells in normal liver.These cells will be useful for the in vitro study of the molecular mechanisms of bile duct development and have important potential for therapeutic strategies, including bioengineered liver approaches.

View Article: PubMed Central - PubMed

Affiliation: INSERM, U972, Paul Brousse Hospital, Villejuif, France; Université Paris Sud, UMR-S 972, Villejuif, France; IFR 93, Bicêtre Hospital, Kremlin-Bicêtre, France; DHU Hepatinov, Paul Brousse Hospital, Villejuif, France.

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Agonists induce Ca2+ increase in hESC-Chol. (A) RT-PCR analysis of gene expression of receptors involved in intracellular Ca2+ signaling: P2RY1, AChR M3, SSTR2, INSP3R type III. (B,C) Fura-2-loaded hESC-Chol were stimulated either with acetylcholine (ACh, 1 μM), somatostatin (1 nM), or ATP (30 μM) for the times indicated by the horizontal bars. Traces have been shifted arbitrarily along the y-axis for clarity. For technical convenience, traces were interrupted during the washes (each gap lasted 5 minutes). The traces shown are representative of the Ca2+ signal observed in the presence of these different agonists in responding cells in four independent experiments. (D) Summary of the Ca2+ induction data (mean ± SEM).
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fig04: Agonists induce Ca2+ increase in hESC-Chol. (A) RT-PCR analysis of gene expression of receptors involved in intracellular Ca2+ signaling: P2RY1, AChR M3, SSTR2, INSP3R type III. (B,C) Fura-2-loaded hESC-Chol were stimulated either with acetylcholine (ACh, 1 μM), somatostatin (1 nM), or ATP (30 μM) for the times indicated by the horizontal bars. Traces have been shifted arbitrarily along the y-axis for clarity. For technical convenience, traces were interrupted during the washes (each gap lasted 5 minutes). The traces shown are representative of the Ca2+ signal observed in the presence of these different agonists in responding cells in four independent experiments. (D) Summary of the Ca2+ induction data (mean ± SEM).

Mentions: RT-PCR on hESC-Chol revealed that the four different receptors were specifically expressed in these cells compared to human hepatocytes (Fig. 4A). ATP, acetylcholine, or somatostatin stimulation resulted in a Ca2+ increase (Fig. 4B). The percentage of responsive cells was quantified to be 70% ± 5%, 40% ± 4%, and 31% ± 9%, respectively (Fig. 4C).


Generation of functional cholangiocyte-like cells from human pluripotent stem cells and HepaRG cells.

Dianat N, Dubois-Pot-Schneider H, Steichen C, Desterke C, Leclerc P, Raveux A, Combettes L, Weber A, Corlu A, Dubart-Kupperschmitt A - Hepatology (2014)

Agonists induce Ca2+ increase in hESC-Chol. (A) RT-PCR analysis of gene expression of receptors involved in intracellular Ca2+ signaling: P2RY1, AChR M3, SSTR2, INSP3R type III. (B,C) Fura-2-loaded hESC-Chol were stimulated either with acetylcholine (ACh, 1 μM), somatostatin (1 nM), or ATP (30 μM) for the times indicated by the horizontal bars. Traces have been shifted arbitrarily along the y-axis for clarity. For technical convenience, traces were interrupted during the washes (each gap lasted 5 minutes). The traces shown are representative of the Ca2+ signal observed in the presence of these different agonists in responding cells in four independent experiments. (D) Summary of the Ca2+ induction data (mean ± SEM).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4315871&req=5

fig04: Agonists induce Ca2+ increase in hESC-Chol. (A) RT-PCR analysis of gene expression of receptors involved in intracellular Ca2+ signaling: P2RY1, AChR M3, SSTR2, INSP3R type III. (B,C) Fura-2-loaded hESC-Chol were stimulated either with acetylcholine (ACh, 1 μM), somatostatin (1 nM), or ATP (30 μM) for the times indicated by the horizontal bars. Traces have been shifted arbitrarily along the y-axis for clarity. For technical convenience, traces were interrupted during the washes (each gap lasted 5 minutes). The traces shown are representative of the Ca2+ signal observed in the presence of these different agonists in responding cells in four independent experiments. (D) Summary of the Ca2+ induction data (mean ± SEM).
Mentions: RT-PCR on hESC-Chol revealed that the four different receptors were specifically expressed in these cells compared to human hepatocytes (Fig. 4A). ATP, acetylcholine, or somatostatin stimulation resulted in a Ca2+ increase (Fig. 4B). The percentage of responsive cells was quantified to be 70% ± 5%, 40% ± 4%, and 31% ± 9%, respectively (Fig. 4C).

Bottom Line: In addition, we showed that cholangiocyte-like cells could also be generated from human induced pluripotent stem cells, demonstrating the efficacy of our approach with stem/progenitor cells of diverse origins.We have developed a robust and efficient method for differentiating pluripotent stem cells into cholangiocyte-like cells, which display structural and functional similarities to bile duct cells in normal liver.These cells will be useful for the in vitro study of the molecular mechanisms of bile duct development and have important potential for therapeutic strategies, including bioengineered liver approaches.

View Article: PubMed Central - PubMed

Affiliation: INSERM, U972, Paul Brousse Hospital, Villejuif, France; Université Paris Sud, UMR-S 972, Villejuif, France; IFR 93, Bicêtre Hospital, Kremlin-Bicêtre, France; DHU Hepatinov, Paul Brousse Hospital, Villejuif, France.

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Related in: MedlinePlus