Characterization of calcium signals in human induced pluripotent stem cell-derived dentate gyrus neuronal progenitors and mature neurons, stably expressing an advanced calcium indicator protein

Gergő Vőfély, Tünde Berecz, Eszter Szabó, Kornélia Szebényi, Edit Hathy, Tamás I. Orbán, Balázs Sarkadi, László Homolya, Maria C. Marchetto, János M. Réthelyi, Ágota Apáti

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Pluripotent stem cell derived human neuronal progenitor cells (hPSC-NPCs) and their mature neuronal cell culture derivatives may efficiently be used for central nervous system (CNS) drug screening, including the investigation of ligand-induced calcium signalization. We have established hippocampal NPC cultures derived from human induced PSCs, which were previously generated by non-integrating Sendai virus reprogramming. Using established protocols these NPCs were differentiated into hippocampal dentate gyrus neurons. In order to study calcium signaling without the need of dye loading, we have stably expressed an advanced calcium indicator protein (GCaMP6fast) in the NPCs using the Sleeping Beauty transposon system. We observed no significant effects of the long-term GCaMP6 expression on NPC morphology, gene expression pattern or neural differentiation capacity. In order to compare the functional properties of GCaMP6-expressing neural cells and the corresponding parental cells loaded with calcium indicator dye Fluo-4, a detailed characterization of calcium signals was performed. We found that the calcium signals induced by ATP, glutamate, LPA, or proteases - were similar in these two systems. Moreover, the presence of the calcium indicator protein allowed for a sensitive, repeatable detection of changes in calcium signaling during the process of neurogenesis and neuronal maturation.

Original languageEnglish
Pages (from-to)222-230
Number of pages9
JournalMolecular and Cellular Neuroscience
Volume88
DOIs
Publication statusPublished - Apr 2018

Keywords

  • Calcium-imaging
  • Dentate gyrus
  • GCaMP6
  • Hippocampus
  • Human iPSC
  • Neuronal differentiation

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience
  • Cell Biology

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