An attempt to generate neurons from an astrocyte progenitor cell line FBD-104.
In the present study, a clonal astrocyte progenitor cell line derived from p53-deficient fetal brains, named FBD-104, was characterized in monolayer and suspension culture. In monolayer culture with medium containing 10% serum, FBD-104 cells expressed some markers of astrocytes, such as glial fibrillary acidic protein (GFAP), S100beta, and glutamate aspartate transporter (GLAST). They never expressed any markers of neurons or oligodendrocytes. Thus the cell line appears to be restricted to the astroglial lineage. However, in suspension culture in serum-free medium supplemented with EGF and FGF2, FBD-104 cells proliferated and formed neurospheres expressing mRNAs for Mash1 and Math3, generating cells expressing neuron specific beta-III tubulin. Re-plating the spheres onto an adhesive substrate and withdrawal of the growth factors induced the expression of mRNAs for NeuroD and Olig2 and generated more beta-III tubulin-positive cells. The present study demonstrated that neurosphere culture is an efficient method to induce neurogenesis from the astrocyte progenitor cell line FBD-104. We also determined that pretreatment with FGF2 caused a significant increase in yield of neurospheres. Thus, the FBD-104 line is an interesting in vitro model to study effect of trophic factors and adhesive substrates on lineage determination of neural progenitor cells.
Horiuchi
Makoto
M
Department of Biological Science and Technology, Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda City, Chiba 278-8510, Japan.
Tomooka
Yasuhiro
Y
D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D015139:Blotting, Southern; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line / Q000166:cytology / Q000378:metabolism; D019070:Cell Lineage / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014162:Transfection
D016222:Fibroblast Growth Factor 2