Activin A and Wnt-dependent specification of human definitive endoderm cells

作者：Toivonen Sanna^*; Lundin Karolina; Balboa Diego; Ustinov Jarkko; Tamminen Kaisa; Palgi Jaan; Trokovic Ras; Tuuri Timo; Otonkoski Timo

来源：Experimental Cell Research, 2013, 319(17): 2535-2544.

DOI：10.1016/j.yexcr.2013.07.007

• Definitive endoderm
• Human pluripotent stem cells
• Differentiation

Activin/Nodal and Wnt signaling are known to play important roles in the regional specification of endoderm. Here we have investigated the effect of the length of stimulation with Activin A plus Wnt3a on the development of hepatic and pancreatic progenitors from the definitive endoderm (DE) cells derived from human pluripotent stem cells (hPSC). We show that DE-cells derived from hPSC with 3 days high Activin A and Wnt3a treatment were able to differentiate further into both tested endodermal lineages. When prolonging the DE-induction protocol from 3 to 5 or 7 days, almost pure DE-marker positive cell populations were obtained. However, these cells had an impaired pancreatic differentiation capacity, while they still developed into hepatocyte-like cells. Further propagation of the DE-cells in the presence of Wnt3a and Activin A led to the complete loss of differentiation capacity into hepatic or pancreatic lineages. When Wnt3a was removed after 24 h from the initiation of the differentiation, the cells were able to differentiate into PDX1 +/NKX6.1+ pancreatic progenitors even with longer DE induction time while efficiency of hepatic differentiation was lower. Our results suggest that both the length and the timing of Wnt3a treatment during DE induction are crucial for the final differentiation outcome. Alt

• 出版日期2013-10-15