Large-scale generation of human ipsc-derived  neural stem cells/early neural progenitor cells and their neuronal differentiation

Leonardo D’Aiuto, Yun Zhi, Dhanjit Kumar Das, Madeleine R. Wilcox, Jon W. Johnson, Lora Mc Clain, Matthew L. Macdonald, Roberto Di Maio, Mark E. Schurdak, Paolo Piazza, Luigi Viggiano, Robert Sweet, Paul R. Kinchington, Ayantika G. Bhattacharjee, Robert Yolken, Vishwajit L. Nimgaonka

Research output: Contribution to journalArticle

Abstract

Induced pluripotent stem cell (iPSC)-based technologies offer an unprecedented opportunity to perform highthroughput screening of novel drugs for neurological and neurodegenerative diseases. Such screenings require a robust and scalable method for generating large numbers of mature, differentiated neuronal cells. Currently available methods based on differentiation of embryoid bodies (EBs) or directed differentiation of adherent culture systems are either expensive or are not scalable. We developed a protocol for large-scale generation of neuronal stem cells (NSCs)/early neural progenitor cells (eNPCs) and their differentiation into neurons. Our scalable protocol allows robust and cost-effective generation of NSCs/eNPCs from iPSCs. Following culture in neurobasal medium supplemented with B27 and BDNF, NSCs/ eNPCs differentiate predominantly into vesicular glutamate transporter 1 (VGLUT1) positive neurons. Targeted mass spectrometry analysis demonstrates that iPSC-derived neurons express ligand-gated channels and other synaptic proteins and whole-cell patch-clamp experiments indicate that these channels are functional. The robust and cost-effective differentiation protocol described here for large-scale generation of NSCs/eNPCs and their differentiation into neuronspaves the way for automated high-throughput screening of drugs for neurological and neurodegenerative diseases.

Original languageEnglish (US)
Pages (from-to)365-377
Number of pages13
JournalOrganogenesis
Volume10
Issue number4
DOIs
StatePublished - 2014

Keywords

  • Induced pluripotent stem cells (iPSCs)
  • Neural progenitor cells
  • Neural stem cells
  • Neuronal differentiation
  •  high-throughput screening

ASJC Scopus subject areas

  • Biomedical Engineering
  • Embryology
  • Developmental Biology
  • Transplantation

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  • Cite this

    D’Aiuto, L., Zhi, Y., Kumar Das, D., Wilcox, M. R., Johnson, J. W., Mc Clain, L., Macdonald, M. L., Di Maio, R., Schurdak, M. E., Piazza, P., Viggiano, L., Sweet, R., Kinchington, P. R., Bhattacharjee, A. G., Yolken, R., & Nimgaonka, V. L. (2014). Large-scale generation of human ipsc-derived  neural stem cells/early neural progenitor cells and their neuronal differentiation. Organogenesis, 10(4), 365-377. https://doi.org/10.1080/15476278.2015.1011921