Stem Cell Reviews and Reports

, Volume 6, Issue 2, pp 270–281 | Cite as

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

  • Dae-Sung Kim
  • Jae Souk Lee
  • Joong Woo Leem
  • Yong Jun Huh
  • Ji Young Kim
  • Han-Soo Kim
  • In-Hyun Park
  • George Q. Daley
  • Dong-Youn Hwang
  • Dong-Wook Kim
Article

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.

Keywords

Pluripotent stem cell Differentiation propensity Neural induction Cell signaling Small molecule 

Supplementary material

12015_2010_9138_MOESM1_ESM.ppt (480 kb)
Supplementary Fig. 1NPCs 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)
12015_2010_9138_MOESM2_ESM.ppt (104 kb)
Supplementary Table 1PCR primer sequences used in this study (PPT 103 kb)

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Dae-Sung Kim
    • 1
  • Jae Souk Lee
    • 1
  • Joong Woo Leem
    • 1
  • Yong Jun Huh
    • 1
  • Ji Young Kim
    • 1
  • Han-Soo Kim
    • 2
  • In-Hyun Park
    • 3
  • George Q. Daley
    • 3
  • Dong-Youn Hwang
    • 4
  • Dong-Wook Kim
    • 1
    • 5
  1. 1.Department of PhysiologyBrain Korea 21 Project for Medical ScienceSeoulKorea
  2. 2.Department of Laboratory MedicineYonsei University College of MedicineSeoulKorea
  3. 3.Department of Medicine, Division of Pediatric Hematology Oncology, Children’s Hospital Boston, and Dana-Farber Cancer Institute; Department of Biological Chemistry and Molecular PharmacologyHarvard Medical SchoolBostonUSA
  4. 4.CHA Stem Cell InstituteCHA University College of MedicineSeoulKorea
  5. 5.Department of Physiology, Center for Cell TherapyYonsei University College of MedicineSeoulKorea

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