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Neurotrophins mediate human embryonic stem cell survival

Nature Biotechnology volume 24, pages 344–350 (2006)Cite this article

Abstract

Growth of human embryonic stem (hES) cells as a pluripotent population requires a balance between survival, proliferation and self-renewal signals. Here we demonstrate that hES cells express receptors of the tropomyosin-related kinase (TRK) family, which mediate antiapoptotic signals. We show that three TRK ligands, brain-derived neurotrophic factor, neurotrophin 3 and neurotrophin 4, are survival factors for hES cells. Addition of neurotrophins to hES cell cultures effects a 36-fold improvement in their clonal survival. hES cell cultures maintained in medium containing neurotrophins remain diploid and retain full developmental potency. In the presence of neurotrophins, TRK receptors in hES cells are phosphorylated; TRK receptor inhibition leads to hES cell apoptosis. The survival activity of neurotrophins in hES cells is mediated by the phosphatidylinositol-3-kinase pathway but not the mitogen-activated protein kinase pathway. Neurotrophins improve hES cell survival and may facilitate their manipulation and the development of high-throughput screens to identify factors responsible for hES cell differentiation.

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Figure 1: Human embryonic stem (hES) cells express members of the TRK receptor tyrosine kinase family of neurotrophin receptors.
Figure 2: Neurotrophins are expressed by MEFs.
Figure 3: Disruption of neurotrophin signaling leads to TRK receptor dephosphorylation and hES cell death.
Figure 4: PI-3K activity is required for hES cell survival.

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Acknowledgements

We are indebted to Linzhao Cheng, Ira Daar, Lino Tessarollo, Brad Harris, John Gearhart and Ian McNiece for helpful suggestions and comments; Lino Tessarollo, Gautam Dravid, David Ginty, Hongjun Song, Xin Duan, Pantelis Tsoulfas, Karen Lackey and Rasi Wickramsinghe for reagents and advice; and Gail Stetten and the Cytogenetics Facility for karyotype analysis. We are also grateful to Sean Donovan and Robert Clouse III for their support and patience and members of the Donovan Lab for their support and encouragement. L.F.L. and P.J.D. dedicate this work to Camilynn I. Brannan (1963–2002). This work was supported by a National Research Service Award (to A.D.P.) by a National Institutes of Health Grant (to P.J.D.) and by institutional funds from the Johns Hopkins University School of Medicine (to L.F.L. and P.J.D.).

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Author notes

  1. Leslie F Lock & Peter J Donovan

    Present address: Stem Cell Program and Departments of Developmental and Cell Biology and of Biological Chemistry, Hewitt Hall, University of California, Irvine, Irvine, California, 92697, USA

Authors and Affiliations

  1. Germ Cell and Stem Cell Group, Stem Cell Biology Program, Institute for Cell Engineering, Johns Hopkins University, School of Medicine, Broadway Research Building, Suite 759, 733 North Broadway, Baltimore, 21205, Maryland, USA

    April D Pyle & Peter J Donovan

  2. Genetic Models Group, Stem Cell Biology Program, Institute for Cell Engineering, Johns Hopkins University, School of Medicine, Broadway Research Building, Suite 759, 733 North Broadway, Baltimore, 21205, Maryland, USA

    Leslie F Lock

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Correspondence to Peter J Donovan.

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Supplementary information

Supplementary Fig. 1

Expression of neurotrophins receptors on H9 hES cells and controls for antibody reactivity. (PDF 701 kb)

Supplementary Fig. 2

Effect of neurotrophins on hES cell differentiation and TRK phosphorylation. (PDF 1602 kb)

Supplementary Fig. 3

Effect of neurotrophins on the population doubling time of hES cells. (PDF 875 kb)

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Pyle, A., Lock, L. & Donovan, P. Neurotrophins mediate human embryonic stem cell survival. Nat Biotechnol 24, 344–350 (2006). https://doi.org/10.1038/nbt1189

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