Study on cell lineage of neuronal stem cell in vitro

Research Project

Project/Area Number	14580778
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Neuroscience in general
Research Institution	Soka University
Principal Investigator	YAMANOHA Banri Soka University, Department of Environmental Engineering for Symbiosis, Faculty of Engineering, 工学部環境共生工学科, 講師 (60247286)
Project Period (FY)	2002 – 2004
Project Status	Completed (Fiscal Year 2004)
Budget Amount *help	¥3,500,000 (Direct Cost: ¥3,500,000) Fiscal Year 2004: ¥100,000 (Direct Cost: ¥100,000) Fiscal Year 2003: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 2002: ¥3,100,000 (Direct Cost: ¥3,100,000)
Keywords	Neural stem cells / Neural precursor cell / Flow cytometry / Primary culture / basic FGF / Neurospheres / EGF / Differentiation / 前駆細胞 / Neurospheres / bFGF
Research Abstract	Basic fibroblast growth factor(bFGF) and epidermal growth factor(EGF) stimulates the proliferation of neural stem cells from early developmental brain. For the receptor of the growth factor on the cells, the timing of expression in culture system is seem to affects its cell lineage. In this study, focus on cell lineage of neural stem cells. The cells are separated from forebrain in embryonic stage at 12 day. The cell isolation method is established to use flow cytometry with light scattering. Isolation qualify in this method would be highly to purify neural stem cells from the cell suspension which effects to cellular events in culture system. In cell suspension of forebrain, A2B5 bound cells were appeared 12% in total cells, whereas almost cells were belonged to nestin-positive cells. The bound cells in both antibody were also appeared in the suspension. Cell culture is performed in differentiation medium in cells isolated by flow cytometry, respectively. These cell populations were generate neuronal cells first and glial cells later. The cells generate neural progenitor and glial progenitor cells randomly. This result would seem that glial progenitor was not accepted FGF signaling in culture period at 8 to 10 days. In medium containing EGF, neural stem cells were appeared neurospheres at 8 to 10 days, whereas not before 8 day. It seem to be a different between FGF and EGF stimulate cells.