Requirement for neurogenesis to proceed through the division of neuronal progenitors following differentiation of epidermal growth factor and fibroblast growth factor-2-responsive human neural stem cells

被引:88
作者
Ostenfeld, T
Svendsen, CN
机构
[1] Univ Wisconsin, Waisman Ctr, Stem Cell Res Program, Dept Anat, Madison, WI 53705 USA
[2] Univ Wisconsin, Waisman Ctr, Stem Cell Res Program, Dept Neurol, Madison, WI 53705 USA
[3] Univ Cambridge, Ctr Brain Repair, Cambridge CB2 1TN, England
关键词
human stem cell; mitogen; neurogenesis; progenitor; epidermal growth factor; fibroblast growth factor-2;
D O I
10.1634/stemcells.22-5-798
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Epidermal growth factor (EGF)- and fibroblast growth factor-2 (FGF-2)-responsive human neural stem cells may provide insight into mechanisms of neural development and have applications in cell-based therapeutics for neurological disease. However, their biology after expansion in vitro is currently poorly understood. Cells grown in either EGF or FGF-2 or a combination of both mitogens displayed characteristically similar levels of transcriptional activation and comparable proliferative profiles with linear cell-cycle kinetics and possessed similar neuronal differentiation capabilities. These data support the view that human neurospheres at later stages of expansion (>10 weeks) are comprised overwhelmingly of a single type of stem cell responsive to both EGF and FGF-2. After mitogen withdrawal and neurosphere plating, bromodeoxyuridine pulse-chase experiments revealed that the stem cells did not undergo differentiation directly into neurons. Instead, most immature neurons arose via the division of emerging progenitor cells in the absence of exogenous EGF or FGF-2. Neurogenesis was abolished by application of high concentrations of either EGF/FGF-2 or the mitotic inhibitor cytosine-b-arabinofuranoside, suggesting that there is an obligatory requirement for at least one round of cell division in the absence of mitogens as a prelude to terminal neuronal differentiation. The differentiation of human neurospheres provides a useful model of human neurogenesis, and the data presented indicate that it proceeds through the division of committed neuronal progenitor cells rather than directly from the neural stem cell.
引用
收藏
页码:798 / 811
页数:14
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