Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells

C H.-H Cho, N. R.-F Hannan, F. M. Docherty, H. M. Docherty, Maria Joao Marques de Lima, M. W. B. Trotter, K. Docherty, L. Vallier

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Abstract

Abstract
Aims/hypothesis
Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hIPSCs) offer unique opportunities for regenerative medicine and for the study of mammalian development. However, developing methods to differentiate hESCs/hIPSCs into specific cell types following a natural pathway of development remains a major challenge.
Methods
We used defined culture media to identify signalling pathways controlling the differentiation of hESCs/hIPSCs into pancreatic or hepatic progenitors. This approach avoids the use of feeders, stroma cells or serum, all of which can interfere with experimental outcomes and could preclude future clinical applications.
Results
This study reveals, for the first time, that activin/TGF-ß signalling blocks pancreatic specification induced by retinoic acid while promoting hepatic specification in combination with bone morphogenetic protein and fibroblast growth factor. Using this knowledge, we developed culture systems to differentiate human pluripotent stem cells into near homogenous population of pancreatic and hepatic progenitors displaying functional characteristics specific to their natural counterparts. Finally, functional experiments showed that activin/TGF-ß signalling achieves this essential function by controlling the levels of transcription factors necessary for liver and pancreatic development, such as HEX and HLXB9.
Conclusion/interpretation
Our methods of differentiation provide an advantageous system to model early human endoderm development in vitro, and also represent an important step towards the generation of pancreatic and hepatic cells for clinical applications.
Original languageEnglish
Pages (from-to)3284-3295
Number of pages12
JournalDiabetologia
Volume55
Issue number12
Early online date26 Sep 2012
DOIs
Publication statusPublished - Dec 2012

Fingerprint

Activins
Pluripotent Stem Cells
Induced Pluripotent Stem Cells
Liver
Feeder Cells
Endoderm
Bone Morphogenetic Proteins
Regenerative Medicine
Fibroblast Growth Factors
Human Development
Tretinoin
Culture Media
Hepatocytes
Transcription Factors
Inhibition (Psychology)
Serum
Population
Human Embryonic Stem Cells

Keywords

  • activin/nodal signalling
  • beta cell
  • hepatocyte
  • human pluripotent stem cells
  • liver
  • pancreas
  • PDX1
  • TGF-ß

Cite this

Cho, C. H. -H., Hannan, N. R. -F., Docherty, F. M., Docherty, H. M., Marques de Lima, M. J., Trotter, M. W. B., ... Vallier, L. (2012). Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells. Diabetologia, 55(12), 3284-3295. https://doi.org/10.1007/s00125-012-2687-x

Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells. / Cho, C H.-H; Hannan, N. R.-F; Docherty, F. M.; Docherty, H. M.; Marques de Lima, Maria Joao; Trotter, M. W. B. ; Docherty, K.; Vallier, L.

In: Diabetologia, Vol. 55, No. 12, 12.2012, p. 3284-3295.

Research output: Contribution to journalArticle

Cho, CH-H, Hannan, NR-F, Docherty, FM, Docherty, HM, Marques de Lima, MJ, Trotter, MWB, Docherty, K & Vallier, L 2012, 'Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells', Diabetologia, vol. 55, no. 12, pp. 3284-3295. https://doi.org/10.1007/s00125-012-2687-x
Cho CH-H, Hannan NR-F, Docherty FM, Docherty HM, Marques de Lima MJ, Trotter MWB et al. Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells. Diabetologia. 2012 Dec;55(12):3284-3295. https://doi.org/10.1007/s00125-012-2687-x
Cho, C H.-H ; Hannan, N. R.-F ; Docherty, F. M. ; Docherty, H. M. ; Marques de Lima, Maria Joao ; Trotter, M. W. B. ; Docherty, K. ; Vallier, L. / Inhibition of activin/nodal signalling is necessary for pancreatic differentiation of human pluripotent stem cells. In: Diabetologia. 2012 ; Vol. 55, No. 12. pp. 3284-3295.
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abstract = "Abstract Aims/hypothesis Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hIPSCs) offer unique opportunities for regenerative medicine and for the study of mammalian development. However, developing methods to differentiate hESCs/hIPSCs into specific cell types following a natural pathway of development remains a major challenge. Methods We used defined culture media to identify signalling pathways controlling the differentiation of hESCs/hIPSCs into pancreatic or hepatic progenitors. This approach avoids the use of feeders, stroma cells or serum, all of which can interfere with experimental outcomes and could preclude future clinical applications. Results This study reveals, for the first time, that activin/TGF-{\ss} signalling blocks pancreatic specification induced by retinoic acid while promoting hepatic specification in combination with bone morphogenetic protein and fibroblast growth factor. Using this knowledge, we developed culture systems to differentiate human pluripotent stem cells into near homogenous population of pancreatic and hepatic progenitors displaying functional characteristics specific to their natural counterparts. Finally, functional experiments showed that activin/TGF-{\ss} signalling achieves this essential function by controlling the levels of transcription factors necessary for liver and pancreatic development, such as HEX and HLXB9. Conclusion/interpretation Our methods of differentiation provide an advantageous system to model early human endoderm development in vitro, and also represent an important step towards the generation of pancreatic and hepatic cells for clinical applications.",
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AU - Marques de Lima, Maria Joao

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AB - Abstract Aims/hypothesis Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hIPSCs) offer unique opportunities for regenerative medicine and for the study of mammalian development. However, developing methods to differentiate hESCs/hIPSCs into specific cell types following a natural pathway of development remains a major challenge. Methods We used defined culture media to identify signalling pathways controlling the differentiation of hESCs/hIPSCs into pancreatic or hepatic progenitors. This approach avoids the use of feeders, stroma cells or serum, all of which can interfere with experimental outcomes and could preclude future clinical applications. Results This study reveals, for the first time, that activin/TGF-ß signalling blocks pancreatic specification induced by retinoic acid while promoting hepatic specification in combination with bone morphogenetic protein and fibroblast growth factor. Using this knowledge, we developed culture systems to differentiate human pluripotent stem cells into near homogenous population of pancreatic and hepatic progenitors displaying functional characteristics specific to their natural counterparts. Finally, functional experiments showed that activin/TGF-ß signalling achieves this essential function by controlling the levels of transcription factors necessary for liver and pancreatic development, such as HEX and HLXB9. Conclusion/interpretation Our methods of differentiation provide an advantageous system to model early human endoderm development in vitro, and also represent an important step towards the generation of pancreatic and hepatic cells for clinical applications.

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