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Robust Enhancement of Neural Differentiation from Human ES and iPS Cells Regardless of their Innate Difference in Differentiation Propensity

Stem Cell Reviews and Reports volume 6, pages 270–281 (2010)Cite this article

An Erratum to this article was published on 23 April 2010

Abstract

Our analyses of three human induced pluripotent stem cell (hiPSC) and six human embryonic stem cell (hESC) lines showed marked variability in differentiation potential into specific lineages, which often hampers their differentiation into specific cell types or cell lineages of interest. Simultaneous inhibition of both Activin/Nodal and BMP pathways with small molecules, SB431542 and dorsomorphin (DM), respectively, promoted significant neural differentiation from all human pluripotent stem cell (hPSC) lines tested, regardless of their differentiation propensity. On the contrary, differentiation into other cell lineages and the number of undifferentiated cells were significantly reduced after differentiation by the dual inhibition. These results demonstrate that innate differentiation propensity of hPSCs could be overcome, at least in part, by modulation of intracellular signaling pathways, resulting in efficient generation of desirable cell types, such as neural cells.

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References

  1. Osafune, K., Caron, L., Borowiak, M., et al. (2008). Marked differences in differentiation propensity among human embryonic stem cell lines. Nature Biotechnology, 26, 313–315.

    Article  CAS  PubMed  Google Scholar 

  2. Oh, S. K., Kim, H. S., Park, Y. B., et al. (2005). Methods for expansion of human embryonic stem cells. Stem Cells, 23, 605–609.

    Article  CAS  PubMed  Google Scholar 

  3. Park, I. H., Zhao, R., West, J. A., et al. (2007). Reprogramming of human somatic cells to pluripotency with defined factors. Nature, 451, 141–146.

    Article  PubMed  Google Scholar 

  4. Cho, M. S., Lee, Y. E., Kim, J. Y., et al. (2008). Highly efficient and large-scale generation of functional dopamine neurons from human embryonic stem cells. Proceedings of the National Academy of Sciences of the United States of America, 105, 3392–3397.

    Article  CAS  PubMed  Google Scholar 

  5. Zhang, S. C., Wernig, M., Duncan, I. D., et al. (2001). In vitro differentiation of transplantable neural precursors from human embryonic stem cells. Nature Biotechnology, 19, 1129–1133.

    Article  CAS  PubMed  Google Scholar 

  6. Pfaffl, M. W. (2001). In vitro differentiation of transplantable neural precursors from human embryonic stem cells. Nucleic Acids Research, 29, e45.

    Article  CAS  PubMed  Google Scholar 

  7. Wilson, S. I., & Edlund, T. (2001). Neural induction: toward a unifying mechanism. Nature Neuroscience, 4(Suppl), 1161–1168.

    Article  CAS  PubMed  Google Scholar 

  8. Muñoz-Sanjuán, I., & Brivanlou, A. H. (2002). Neural induction, the default model and embryonic stem cells. Nature Reviews Neuroscience, 3, 271–280.

    Article  PubMed  Google Scholar 

  9. Yu, P. B., Hong, C. C., Sachidanandan, C., et al. (2008). Dorsomorphin inhibits BMP signals required for embryogenesis and iron metabolism. Nature Chemical Biology, 4, 33–41.

    Article  CAS  PubMed  Google Scholar 

  10. Schier, A. F. (2003). Nodal signaling in vertebrate development. Annual Review of Cell and Developmental Biology, 19, 589–621.

    Article  CAS  PubMed  Google Scholar 

  11. Vallier, L., Reynolds, D., & Pedersen, R. A. (2004). Nodal inhibits differentiation of human embryonic stem cells along the neuroectodermal default pathway. Developmental Biology, 275, 403–421.

    Article  CAS  PubMed  Google Scholar 

  12. Camus, A., Perea-Gomez, A., Moreau, A., et al. (2006). Absence of Nodal signaling promotes precocious neural differentiation in the mouse embryo. Developmental Biology, 295, 743–755.

    Article  CAS  PubMed  Google Scholar 

  13. Vallier, L., Alexander, M., & Pedersen, R. A. (2005). Activin/Nodal and FGF pathways cooperate to maintain pluripotency of human embryonic stem cells. Journal of Cell Science, 118, 4495–4509.

    Article  CAS  PubMed  Google Scholar 

  14. Xiao, L., Yuan, X., & Sharkis, S. J. (2006). Activin A maintains self-renewal and regulates fibroblast growth factor, Wnt, and bone morphogenic protein pathways in human embryonic stem cells. Stem Cells, 24, 1476–1486.

    Article  CAS  PubMed  Google Scholar 

  15. Inman, G. J., Nicolás, F. J., Callahan, J. F., et al. (2002). SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Molecular Pharmacology, 62, 65–74.

    Article  CAS  PubMed  Google Scholar 

  16. Wu, Z., Zhang, W., Chen, G., et al. (2008). Combinatorial signals of activin/nodal and bone morphogenic protein regulate the early lineage segregation of human embryonic stem cells. Journal of Biological Chemistry, 283, 24991–25002.

    Article  CAS  PubMed  Google Scholar 

  17. Chang, C., & Harland, R. M. (2007). Neural induction requires continued suppression of both Smad1 and Smad2 signals during gastrulation. Development, 134, 3861–3872.

    Article  CAS  PubMed  Google Scholar 

  18. Xu, R. H., Chen, X., Li, D. S., et al. (2007). BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nature Biotechnology, 20, 1261–1264.

    Article  Google Scholar 

  19. Pera, M. F., Andrade, J., Houssami, S., et al. (2004). Regulation of human embryonic stem cell differentiation by BMP-2 and its antagonist noggin. Journal of Cell Science, 117, 1269–1280.

    Article  CAS  PubMed  Google Scholar 

  20. Itsykson, P., Ilouz, N., Turetsky, T., et al. (2005). Derivation of neural precursors from human embryonic stem cells in the presence of noggin. Molecular and Cellular Neurosciences, 30, 24–36.

    Article  CAS  PubMed  Google Scholar 

  21. Shen, M. M. (2007). Nodal signaling: developmental roles and regulation. Development, 134, 1023–1034.

    Article  CAS  PubMed  Google Scholar 

  22. James, D., Levine, A. J., Besser, D., et al. (2005). TGFbeta/activin/nodal signaling is necessary for the maintenance of pluripotency in human embryonic stem cells. Development, 132, 1273–1282.

    Article  CAS  PubMed  Google Scholar 

  23. Beattie, G. M., Lopez, A. D., Bucay, N., et al. (2005). Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. Stem Cells, 23, 489–495.

    Article  CAS  PubMed  Google Scholar 

  24. Xu, R. H., Peck, R. M., Li, D. S., et al. (2005). Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells. Nature Methods, 2, 185–90.

    Article  CAS  PubMed  Google Scholar 

  25. Chambers, S. M., Fasano, C. A., Papapetrou, E. P., et al. (2009). Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature Biotechnology, 27, 275–280.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgement

This research was supported by grants (code: SC1110, SC4140 and SC5170) from the Stem Cell Research Center of the 21th Century Frontier Research Program funded by the Ministry of Education, Science and Technology, Republic of Korea.

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Authors and Affiliations

  1. Department of Physiology, Brain Korea 21 Project for Medical Science, Seoul, 120-752, Korea

    Dae-Sung Kim, Jae Souk Lee, Joong Woo Leem, Yong Jun Huh, Ji Young Kim & Dong-Wook Kim

  2. Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, 120-752, Korea

    Han-Soo Kim

  3. Department of Medicine, Division of Pediatric Hematology Oncology, Children’s Hospital Boston, and Dana-Farber Cancer Institute; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Karp Family Research Building 7214, 300 Longwood Avenue, Boston, MA, 02115, USA

    In-Hyun Park & George Q. Daley

  4. CHA Stem Cell Institute, CHA University College of Medicine, 606-16 Vision bldg., 4th floor, Yeoksam-Dong, Kangnam-Gu, Seoul, Korea

    Dong-Youn Hwang

  5. Department of Physiology, Center for Cell Therapy, Yonsei University College of Medicine, 134 Shichon-dong, Seodaemoon-gu, Seoul, Korea

    Dong-Wook Kim

Authors
  1. Dae-Sung Kim
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  2. Jae Souk Lee
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  4. Yong Jun Huh
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  5. Ji Young Kim
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  6. Han-Soo Kim
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  7. In-Hyun Park
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Corresponding authors

Correspondence to Dong-Youn Hwang or Dong-Wook Kim.

Additional information

Authors contributions:

D-.S.K.: Conception and design, Collection and/or assembly of data, Data analysis and interpretation, Manuscript writing

J. S. L.: Collection and/or assembly of data, Data analysis and interpretation

J.W.L.: Data analysis and interpretation

J.Y.K.: Collection and/or assembly of data, Data analysis and interpretation

Y.J.H.: Collection and/or assembly of data, Data analysis and interpretation

H-.S.K.: Data analysis and interpretation, Manuscript writing

D-.Y.H.: Data analysis and interpretation, Manuscript writing

I-.H.P.: Provision of study materials, Data analysis and interpretation

G.Q.D.: Provision of study materials, Data analysis and interpretation

D-.W.K.: Conception and design, Financial support, Data analysis and interpretation, Manuscript writing and Final approval of manuscript

An erratum to this article can be found at http://dx.doi.org/10.1007/s12015-010-9151-4

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Fig. 1

NPCs derived from hPSCs by simultaneous inhibition of BMP and Activin/Nodal signaling can differentiate into all neural lineages and also into dopaminergic neurons. (A-B) H9-NPCs were generated in the presence of DM and SB431542, followed by expansion in N2 medium supplemented with bFGF. The expanded NPCs were then triturated and grown attached on Matrigel-coated cover-slips in N2 medium supplemented with 2% FBS (Invitrogen) for additional 4 weeks. All three neural cell types, neurons (Tuj1 positive), astrocytes (GFAP positive) (A) and oligodendrocytes (O4 positive) (B), were detected after differentiation. (C) A number of TH-positive neurons were detected when NPCs derived from H9 were subjected to DA differentiation. (PPT 480 kb)

Supplementary Table 1

PCR primer sequences used in this study (PPT 103 kb)

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Kim, DS., Lee, J.S., Leem, J.W. et al. Robust Enhancement of Neural Differentiation from Human ES and iPS Cells Regardless of their Innate Difference in Differentiation Propensity. Stem Cell Rev and Rep 6, 270–281 (2010). https://doi.org/10.1007/s12015-010-9138-1

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  • DOI: https://doi.org/10.1007/s12015-010-9138-1

Keywords

  • Pluripotent stem cell
  • Differentiation propensity
  • Neural induction
  • Cell signaling
  • Small molecule