title: A rapid and efficient method for neuronal induction of the P19 embryonic carcinoma cell line.
abstract: BACKGROUND
  P19 mouse embryonic carcinoma cells are conventionally induced to differentiate into neural cells by suspension culture in the presence of retinoic acid to form cell aggregates, followed by adhesion culture in a poly-l-lysine-coated dish. Drawbacks of this procedure include it taking more than 10 days to obtain mature neurons, and non-neuronal proliferating cells occupying the majority of the cell population with time.
  
  
  METHODS
  Here, we show a novel method for the rapid and efficient neurogenesis of P19 cells, without aggregate formation in a suspension culture. The new approach is based on an adherent serum-free culture in a laminin-coated dish in the presence of FGF8, a γ-secretase inhibitor, and cytosine arabinoside.
  
  
  RESULTS
  The new method efficiently induced P19 cells to differentiate into neurons within 4 days, and subsequently into mature neurons that were responsive to several neurotransmitters, giving spontaneous neuronal network activity within 6 days.
  
  
  CONCLUSIONS
  The novel method accelerated neuritogenesis and enhanced population of neuron selectively compared to the conventional method. Proliferating non-neuronal cells were eliminated by adding cytosine arabinoside during neuronal maturation.
  
  
  CONCLUSIONS
  The method is useful for studying neuronal differentiation or activities.
authors.affiliation: Laboratory of Neuroglycobiology, Department of Molecular Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-motoyama, Kita-ku, Kyoto 603-8555, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Nakayama
authors.affiliation: Laboratory of Neuroglycobiology, Department of Molecular Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-motoyama, Kita-ku, Kyoto 603-8555, Japan.
authors.forename: Ayumi
authors.initials: A
authors.lastname: Wada
authors.affiliation: Laboratory of Neuroglycobiology, Department of Molecular Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-motoyama, Kita-ku, Kyoto 603-8555, Japan.
authors.forename: Rei
authors.initials: R
authors.lastname: Inoue
authors.affiliation: Center for Genomics Medicine, Kyoto University Graduate School of Medicine, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Terasawa
authors.affiliation: Department of Pharmacogenomics, Kyoto University Graduate School of Pharmaceutical Science, Sakyo-ku, Kyoto 606-8501, Japan; Department of Applied Biological Science, Tokyo University of Agriculture and Technology, Fuchu-city, Tokyo 183-8538, Japan.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kimura
authors.affiliation: Laboratory of Neuroglycobiology, Department of Molecular Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-motoyama, Kita-ku, Kyoto 603-8555, Japan.
authors.forename: Naosuke
authors.initials: N
authors.lastname: Nakamura
authors.affiliation: Laboratory of Neuroglycobiology, Department of Molecular Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-motoyama, Kita-ku, Kyoto 603-8555, Japan. Electronic address: kurosaka@cc.kyoto-su.ac.jp.
authors.forename: Akira
authors.initials: A
authors.lastname: Kurosaka
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D018236:Carcinoma, Embryonal / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D045744:Cell Line, Tumor; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D051524:Fibroblast Growth Factor 8 / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000502:physiology; D007797:Laminin / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D055495:Neurogenesis / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D018377:Neurotransmitter Agents / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D013997:Time Factors
chemical_list: D016895:Culture Media, Serum-Free; C496457:Fgf8 protein, mouse; D007797:Laminin; D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; D051524:Fibroblast Growth Factor 8; D011189:Potassium Chloride; D002118:Calcium
keywords: Neural differentiation; Neurotransmitter; P19 cells