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Differentiation of V2a interneurons from human pluripotent stem cells.
Butts, Jessica C; McCreedy, Dylan A; Martinez-Vargas, Jorge Alexis; Mendoza-Camacho, Frederico N; Hookway, Tracy A; Gifford, Casey A; Taneja, Praveen; Noble-Haeusslein, Linda; McDevitt, Todd C.
Afiliação
  • Butts JC; Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158.
  • McCreedy DA; Graduate Program in Bioengineering, University of California, San Francisco, CA 94158.
  • Martinez-Vargas JA; Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158.
  • Mendoza-Camacho FN; Department of Neurosurgery, University of California, San Francisco, CA 94143.
  • Hookway TA; Department of Physical Therapy and Rehabilitation Science, University of California, San Francisco, CA 94143.
  • Gifford CA; Department of Bioengineering, University of California, Berkeley, CA 94709.
  • Taneja P; Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158.
  • Noble-Haeusslein L; Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158.
  • McDevitt TC; Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158.
Proc Natl Acad Sci U S A ; 114(19): 4969-4974, 2017 05 09.
Article em En | MEDLINE | ID: mdl-28438991
The spinal cord consists of multiple neuronal cell types that are critical to motor control and arise from distinct progenitor domains in the developing neural tube. Excitatory V2a interneurons in particular are an integral component of central pattern generators that control respiration and locomotion; however, the lack of a robust source of human V2a interneurons limits the ability to molecularly profile these cells and examine their therapeutic potential to treat spinal cord injury (SCI). Here, we report the directed differentiation of CHX10+ V2a interneurons from human pluripotent stem cells (hPSCs). Signaling pathways (retinoic acid, sonic hedgehog, and Notch) that pattern the neural tube were sequentially perturbed to identify an optimized combination of small molecules that yielded ∼25% CHX10+ cells in four hPSC lines. Differentiated cultures expressed much higher levels of V2a phenotypic markers (CHX10 and SOX14) than other neural lineage markers. Over time, CHX10+ cells expressed neuronal markers [neurofilament, NeuN, and vesicular glutamate transporter 2 (VGlut2)], and cultures exhibited increased action potential frequency. Single-cell RNAseq analysis confirmed CHX10+ cells within the differentiated population, which consisted primarily of neurons with some glial and neural progenitor cells. At 2 wk after transplantation into the spinal cord of mice, hPSC-derived V2a cultures survived at the site of injection, coexpressed NeuN and VGlut2, extended neurites >5 mm, and formed putative synapses with host neurons. These results provide a description of V2a interneurons differentiated from hPSCs that may be used to model central nervous system development and serve as a potential cell therapy for SCI.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Antígenos de Diferenciação / Diferenciação Celular / Células-Tronco Pluripotentes Induzidas / Células-Tronco Embrionárias Humanas / Neurônios Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Antígenos de Diferenciação / Diferenciação Celular / Células-Tronco Pluripotentes Induzidas / Células-Tronco Embrionárias Humanas / Neurônios Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article