Stromal factors SDF1α, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells

Catherine M. Schwartz, Tahereh Tavakoli, Charmaine Jamias, Sung Soo Park, Stuart Maudsley, Bronwen Martin, Terry M. Phillips, Pamela J. Yao, Katsuhiko Itoh, Wu Ma, Mahendra S. Rao, Ernest Arenas, Mark P. Mattson

Research output: Contribution to journalArticle

Abstract

Human embryonic stem cell (hESC)-derived dopaminergic (DA) neurons hold potential for treating Parkinson's disease (PD) through cell replacement therapy. Generation of DA neurons from hESCs has been achieved by coculture with the stromal cell line PA6, a source of stromal cell-derived inducing activity (SDIA). However, the factors produced by stromal cells that result in SDIA are largely undefined. We previously reported that medium conditioned by PA6 cells can generate functional DA neurons from NTera2 human embryonal carcinoma stem cells. Here we show that PA6-conditioned medium can induce DA neuronal differentiation in both NTera2 cells and the hESC I6 cell line. To identify the factor(s) responsible for SDIA, we used large-scale microarray analysis of gene expression combined with mass spectrometric analysis of PA6-conditioned medium (CM). The candidate factors, hepatocyte growth factor (HGF), stromal cell-derived factor-1 α (SDF1α), secreted frizzled-related protein 1 (sFRP1), and vascular endothelial growth factor D (VEGFD) were identified, and their concentrations in PA6 CM were established by immunoaffinity capillary electrophoresis. Upon addition of SDF1α, sFRP1, and VEGFD to the culture medium, we observed an increase in the number of cells expressing tyrosine hydroxylase (a marker for DA neurons) and βIII-tubulin (a marker for immature neurons) in both the NTera2 and I6 cell lines. These results indicate that SDF1α, sFRP1, and VEGFD are major components of SDIA and suggest the potential use of these defined factors to elicit DA differentiation of pluripotent human stem cells for therapeutic intervention in PD.

Original languageEnglish (US)
Pages (from-to)1367-1381
Number of pages15
JournalJournal of Neuroscience Research
Volume90
Issue number7
DOIs
StatePublished - Jul 2012

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Vascular Endothelial Growth Factor D
Chemokine CXCL12
Pluripotent Stem Cells
Dopaminergic Neurons
Stromal Cells
Conditioned Culture Medium
Cell Line
Parkinson Disease
Embryonal Carcinoma Stem Cells
Hepatocyte Growth Factor
Capillary Electrophoresis
Tyrosine 3-Monooxygenase
Tubulin
Microarray Analysis
Cell- and Tissue-Based Therapy
Coculture Techniques
Culture Media
frizzled related protein-1
Cell Count
Gene Expression

Keywords

  • Dopaminergic neurons
  • Embryonic stem cells
  • Neuronal differentiation
  • Stromal cell-derived inducing activity

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

Cite this

Schwartz, C. M., Tavakoli, T., Jamias, C., Park, S. S., Maudsley, S., Martin, B., ... Mattson, M. P. (2012). Stromal factors SDF1α, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells. Journal of Neuroscience Research, 90(7), 1367-1381. https://doi.org/10.1002/jnr.23064

Stromal factors SDF1α, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells. / Schwartz, Catherine M.; Tavakoli, Tahereh; Jamias, Charmaine; Park, Sung Soo; Maudsley, Stuart; Martin, Bronwen; Phillips, Terry M.; Yao, Pamela J.; Itoh, Katsuhiko; Ma, Wu; Rao, Mahendra S.; Arenas, Ernest; Mattson, Mark P.

In: Journal of Neuroscience Research, Vol. 90, No. 7, 07.2012, p. 1367-1381.

Research output: Contribution to journalArticle

Schwartz, CM, Tavakoli, T, Jamias, C, Park, SS, Maudsley, S, Martin, B, Phillips, TM, Yao, PJ, Itoh, K, Ma, W, Rao, MS, Arenas, E & Mattson, MP 2012, 'Stromal factors SDF1α, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells', Journal of Neuroscience Research, vol. 90, no. 7, pp. 1367-1381. https://doi.org/10.1002/jnr.23064
Schwartz, Catherine M. ; Tavakoli, Tahereh ; Jamias, Charmaine ; Park, Sung Soo ; Maudsley, Stuart ; Martin, Bronwen ; Phillips, Terry M. ; Yao, Pamela J. ; Itoh, Katsuhiko ; Ma, Wu ; Rao, Mahendra S. ; Arenas, Ernest ; Mattson, Mark P. / Stromal factors SDF1α, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells. In: Journal of Neuroscience Research. 2012 ; Vol. 90, No. 7. pp. 1367-1381.
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abstract = "Human embryonic stem cell (hESC)-derived dopaminergic (DA) neurons hold potential for treating Parkinson's disease (PD) through cell replacement therapy. Generation of DA neurons from hESCs has been achieved by coculture with the stromal cell line PA6, a source of stromal cell-derived inducing activity (SDIA). However, the factors produced by stromal cells that result in SDIA are largely undefined. We previously reported that medium conditioned by PA6 cells can generate functional DA neurons from NTera2 human embryonal carcinoma stem cells. Here we show that PA6-conditioned medium can induce DA neuronal differentiation in both NTera2 cells and the hESC I6 cell line. To identify the factor(s) responsible for SDIA, we used large-scale microarray analysis of gene expression combined with mass spectrometric analysis of PA6-conditioned medium (CM). The candidate factors, hepatocyte growth factor (HGF), stromal cell-derived factor-1 α (SDF1α), secreted frizzled-related protein 1 (sFRP1), and vascular endothelial growth factor D (VEGFD) were identified, and their concentrations in PA6 CM were established by immunoaffinity capillary electrophoresis. Upon addition of SDF1α, sFRP1, and VEGFD to the culture medium, we observed an increase in the number of cells expressing tyrosine hydroxylase (a marker for DA neurons) and βIII-tubulin (a marker for immature neurons) in both the NTera2 and I6 cell lines. These results indicate that SDF1α, sFRP1, and VEGFD are major components of SDIA and suggest the potential use of these defined factors to elicit DA differentiation of pluripotent human stem cells for therapeutic intervention in PD.",
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AU - Martin, Bronwen

AU - Phillips, Terry M.

AU - Yao, Pamela J.

AU - Itoh, Katsuhiko

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