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

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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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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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  5. Ji Young Kim
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  6. Han-Soo Kim
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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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