A novel three-dimensional culture system for isolation and clonal propagation of neural stem cells using a thermo-reversible gelation polymer

Xin-Zhi Yang, Ken Kataoka, Reinhold Medina, Ken-Ichi Yamamoto, Swe Swe Than, Masahiro Miyazaki, Nam-Ho Huh

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

In the present study, we examined the possible utility of a three-dimensional culture system using a thermo-reversible gelation polymer to isolate and expand neural stem cells (NSCs). The polymer is a synthetic biologically inert polymer and gelates at temperatures higher than the gel-sol transition point ( approximately 20 degrees C). When fetal mouse brain cells were inoculated into the gel, spherical colonies were formed ( approximately 1% in primary culture and approximately 9% in passage cultures). The spheroid-forming cells were positive for expression of the NSC markers nestin and Musashi. Under conditions facilitating spontaneous neural differentiation, the spheroid-forming cells expressed genes characteristic to astrocytes, oligodendrocytes, and neurons. The cells could be successively propagated at least to 80 poly-D-lysines over a period of 20 weeks in the gel culture with a growth rate higher than that observed in suspension culture. The spheroids formed by fetal mouse brain cells in the gel were shown to be of clonal origin. These results indicate that the spheroid culture system is a convenient and powerful tool for isolation and clonal expansion of NSCs in vitro.
Original languageEnglish
Pages (from-to)615-23
Number of pages9
JournalTissue engineering. Part C, Methods
Volume15
Issue number4
DOIs
Publication statusPublished - Dec 2009

Keywords

  • Bromodeoxyuridine
  • Clone Cells
  • Animals
  • Fetus
  • Spheroids, Cellular
  • Temperature
  • Brain
  • Cell Culture Techniques
  • Cell Differentiation
  • Mice
  • Cell Separation
  • Cell Proliferation
  • Nerve Tissue Proteins
  • Gels
  • Neurons
  • Mice, Inbred C57BL
  • Intermediate Filament Proteins
  • Stem Cells
  • Polymers
  • Biological Markers
  • Female

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