title: Characterization of Human Hippocampal Neural Stem/Progenitor Cells and Their Application to Physiologically Relevant Assays for Multiple Ionotropic Glutamate Receptors.
abstract: The hippocampus is an important brain region that is involved in neurological disorders such as Alzheimer disease, schizophrenia, and epilepsy. Ionotropic glutamate receptors-namely,N-methyl-D-aspartate (NMDA) receptors (NMDARs), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors (AMPARs), and kainic acid (KA) receptors (KARs)-are well known to be involved in these diseases by mediating long-term potentiation, excitotoxicity, or both. To predict the therapeutic efficacy and neuronal toxicity of drug candidates acting on these receptors, physiologically relevant systems for assaying brain region-specific human neural cells are necessary. Here, we characterized the functional differentiation of human fetal hippocampus-derived neural stem/progenitor cells-namely, HIP-009 cells. Calcium rise assay demonstrated that, after a 4-week differentiation, the cells responded to NMDA (EC50= 7.5 ± 0.4 µM; n= 4), AMPA (EC50= 2.5 ± 0.1 µM; n= 3), or KA (EC50= 33.5 ± 1.1 µM; n= 3) in a concentration-dependent manner. An AMPA-evoked calcium rise was observed in the absence of the desensitization inhibitor cyclothiazide. In addition, the calcium rise induced by these agonists was inhibited by antagonists for each receptor-namely, MK-801 for NMDA stimulation (IC50= 0.6 ± 0.1 µM; n= 4) and NBQX for AMPA and KA stimulation (IC50= 0.7 ± 0.1 and 0.7 ± 0.03 µM, respectively; n= 3). The gene expression profile of differentiated HIP-009 cells was distinct from that of undifferentiated cells and closely resembled that of the human adult hippocampus. Our results show that HIP-009 cells are a unique tool for obtaining human hippocampal neural cells and are applicable to systems for assay of ionotropic glutamate receptors as a physiologically relevant in vitro model.
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan k7-fukushima@hhc.eisai.co.jp.
authors: Kazuyuki
authors: K
authors: Fukushima
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Yoshikuni
authors: Y
authors: Tabata
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Yoichi
authors: Y
authors: Imaizumi
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Naohiro
authors: N
authors: Kohmura
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Michiko
authors: M
authors: Sugawara
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Kohei
authors: K
authors: Sawada
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Kazuto
authors: K
authors: Yamazaki
authors: Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Masashi
authors: M
authors: Ito
mesh_terms: D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004594:Electrophysiology / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects; D006624:Hippocampus / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007608:Kainic Acid / Q000494:pharmacology; D016202:N-Methylaspartate / Q000494:pharmacology; D058953:Neural Stem Cells / Q000187:drug effects* / Q000378:metabolism; D011810:Quinoxalines / Q000494:pharmacology; D058468:Receptors, Ionotropic Glutamate / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000494:pharmacology
chemical_list: D011810:Quinoxalines; D058468:Receptors, Ionotropic Glutamate; C062865:2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline; D016202:N-Methylaspartate; D016291:Dizocilpine Maleate; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D007608:Kainic Acid; D002118:Calcium
keywords: AMPA receptor; NMDA receptor; human neural stem/progenitor cells; kainic acid receptor; physiologically relevant assay systems