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Culture method for the induction of neurospheres from mouse embryonic stem cells by coculture with PA6 stromal cells.
J Neurosci Res. 2005 May 15; 80(4):467-74.JN

Abstract

Embryonic stem (ES) cells proliferate and maintain their pluripotency for over 1 year in vitro and may therefore provide a sufficient source for cell therapies. However, most of the previously reported methods for obtaining a source for cell therapies have not been simple. We describe here a novel method for induction of neurospheres from mouse ES cells by coculturing on PA6 cells instead of the formation of embryoid bodies. The ES cells cocultured with the PA6 stromal cell line for at least 3 days were capable of differentiating into spheres. The cells in the spheres were all green fluorescent protein (GFP) positive, showing that they were derived from GFP-expressing D3-ES cells. The spheres contained nestin-positive cells. The number of spheres increased when they were cocultured with PA6 for a longer period. Sphere formation was observed even after 10 mechanical dissociations and subculturings, showing its self-renewal ability. The cells differentiated into microtubule-associated protein-2 (MAP2)-positive neuronal cells and glial fibrillary acidic protein (GFAP)-positive glial cells. gamma-Aminobutyric acid-positive cells and tyrosine hydroxylase-positive cells were also observed in the spheres. The percentages of the MAP2- or GFAP-positive cells in the sphere changed according to the period of coculture on PA6 cells. At an early stage of coculture, more neurons were generated and, at a later period, more glial cells were generated. These results suggested that neurosphere could be generated from ES cells by coculturing with PA6, and that these cells resembled neural stem cells derived from mouse fetal brain tissue.

Authors+Show Affiliations

Department of Neurosurgery, Division of Neuroscience, Graduate School of Medicine, Gifu University, Gifu, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

15825193

Citation

Kitajima, Hideomi, et al. "Culture Method for the Induction of Neurospheres From Mouse Embryonic Stem Cells By Coculture With PA6 Stromal Cells." Journal of Neuroscience Research, vol. 80, no. 4, 2005, pp. 467-74.
Kitajima H, Yoshimura S, Kokuzawa J, et al. Culture method for the induction of neurospheres from mouse embryonic stem cells by coculture with PA6 stromal cells. J Neurosci Res. 2005;80(4):467-74.
Kitajima, H., Yoshimura, S., Kokuzawa, J., Kato, M., Iwama, T., Motohashi, T., Kunisada, T., & Sakai, N. (2005). Culture method for the induction of neurospheres from mouse embryonic stem cells by coculture with PA6 stromal cells. Journal of Neuroscience Research, 80(4), 467-74.
Kitajima H, et al. Culture Method for the Induction of Neurospheres From Mouse Embryonic Stem Cells By Coculture With PA6 Stromal Cells. J Neurosci Res. 2005 May 15;80(4):467-74. PubMed PMID: 15825193.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Culture method for the induction of neurospheres from mouse embryonic stem cells by coculture with PA6 stromal cells. AU - Kitajima,Hideomi, AU - Yoshimura,Shinichi, AU - Kokuzawa,Jouji, AU - Kato,Masayasu, AU - Iwama,Toru, AU - Motohashi,Tsutomu, AU - Kunisada,Takahiro, AU - Sakai,Noboru, PY - 2005/4/13/pubmed PY - 2005/7/26/medline PY - 2005/4/13/entrez SP - 467 EP - 74 JF - Journal of neuroscience research JO - J. Neurosci. Res. VL - 80 IS - 4 N2 - Embryonic stem (ES) cells proliferate and maintain their pluripotency for over 1 year in vitro and may therefore provide a sufficient source for cell therapies. However, most of the previously reported methods for obtaining a source for cell therapies have not been simple. We describe here a novel method for induction of neurospheres from mouse ES cells by coculturing on PA6 cells instead of the formation of embryoid bodies. The ES cells cocultured with the PA6 stromal cell line for at least 3 days were capable of differentiating into spheres. The cells in the spheres were all green fluorescent protein (GFP) positive, showing that they were derived from GFP-expressing D3-ES cells. The spheres contained nestin-positive cells. The number of spheres increased when they were cocultured with PA6 for a longer period. Sphere formation was observed even after 10 mechanical dissociations and subculturings, showing its self-renewal ability. The cells differentiated into microtubule-associated protein-2 (MAP2)-positive neuronal cells and glial fibrillary acidic protein (GFAP)-positive glial cells. gamma-Aminobutyric acid-positive cells and tyrosine hydroxylase-positive cells were also observed in the spheres. The percentages of the MAP2- or GFAP-positive cells in the sphere changed according to the period of coculture on PA6 cells. At an early stage of coculture, more neurons were generated and, at a later period, more glial cells were generated. These results suggested that neurosphere could be generated from ES cells by coculturing with PA6, and that these cells resembled neural stem cells derived from mouse fetal brain tissue. SN - 0360-4012 UR - https://www.unboundmedicine.com/medline/citation/15825193/Culture_method_for_the_induction_of_neurospheres_from_mouse_embryonic_stem_cells_by_coculture_with_PA6_stromal_cells_ L2 - https://doi.org/10.1002/jnr.20469 DB - PRIME DP - Unbound Medicine ER -