Medical Molecular Morphology

, Volume 49, Issue 2, pp 119–126 | Cite as

A lower volume culture method for obtaining a larger yield of neuron-like cells from mesenchymal stem cells

  • Atsushi ShimomuraEmail author
  • Akiko Iizuka-Kogo
  • Naoki Yamamoto
  • Ryuji Nomura
Original Paper


Mesenchymal stem cells (MSCs) represent a promising cell source for stem cell therapy to replace neurons damaged by neurodegenerative diseases. A system designed for in vitro neuronal differentiation of MSCs is an indispensable technique, which provides MSC-derived functional neurons for cell-replacement therapies and valuable information in pre-clinical research. This study investigated the effects of reducing the volume of neural induction medium on cell viability and neural differentiation of MSCs. When MSCs were differentiated in low volumes of neural induction medium, rather than using the conventional method, the cell density on culture dishes significantly increased. The % cell death, including apoptosis and necrosis, was significantly lower in the lower volume method than in the conventional method. There were no significant differences between the lower volume and conventional methods in the expression levels of the neuronal marker genes. In an analysis of immunostaining for a mature neuronal marker, no significant difference was detected between the media volumes. These findings demonstrate that neuronal induction of MSCs in low volumes of differentiation medium promoted survival during differentiation and resulted in larger numbers of MSC-derived neurons, compared to the conventional method. This novel lower volume method offers both financial and cell-yield advantages over the conventional method.


A lower volume culture method In vitro neuronal differentiation Mesenchymal stem cell Neural differentiation 



The authors wish to thank Ms. Kazuko Hikita, Mr. Kazuhiro Yanagisawa, Mr. Yohei Takeuchi, Ms. Eri Nishigaki, and Ms. Mayu Tsunekawa for their helpful support. This research was supported by JSPS KAKENHI Grant Number 24592567 and the Aichi Cancer Research Foundation.

Compliance with ethical standards

Conflict of interest

The authors have declared that no competing interests exist.


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Copyright information

© The Japanese Society for Clinical Molecular Morphology 2015

Authors and Affiliations

  • Atsushi Shimomura
    • 1
    • 2
    Email author
  • Akiko Iizuka-Kogo
    • 3
  • Naoki Yamamoto
    • 4
  • Ryuji Nomura
    • 5
  1. 1.Department of Communication DisordersHealth Sciences University of Hokkaido School of Psychological ScienceSapporoJapan
  2. 2.Department of Communication DisordersHealth Sciences University of Hokkaido School of Rehabilitation SciencesTobetsuJapan
  3. 3.Department of Anatomy and Cell BiologyGunma University School of MedicineMaebashiJapan
  4. 4.Laboratory of Molecular Biology and HistochemistryFujita Health University Joint Research LaboratoryToyoakeJapan
  5. 5.Department of Anatomy IFujita Health University School of MedicineToyoakeJapan

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