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 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
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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