Journal
JOURNAL OF NEUROSCIENCE RESEARCH
Volume 86, Issue 1, Pages 35-47Publisher
WILEY
DOI: 10.1002/jnr.21460
Keywords
human neural stem cells; astrocytes; coculture; cell survival; bFGF; cholinergic neurons
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Funding
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [R01NS046025] Funding Source: NIH RePORTER
- NINDS NIH HHS [NS046025] Funding Source: Medline
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Establishment of an in vitro model of human cholinergic neurons would be highly desirable for understanding and developing treatment for Alzheimer's and motoneuron diseases. Previously we reported that the combination of basic fibroblast growth factor (bFGF), heparin, and laminin directs human fetal neural stem cells to form cholinergic neurons. One problem, however, is that long-term in vitro survival of these cells is low. Our goal for this study was to determine whether astrocytes or their secreted factors enhance differentiation and survival of cholinergic neurons under long-term differentiation conditions. We demonstrate here that astrocytes or astrocyte conditioned media did not enhance cholinergic differentiation but did increase the long-term survival of differentiated human neural stem cells, particularly cholinergic neurons. We further show that astrocytes protected long-term-differentiated cells from apoptotic cell death, which is at least partially mediated by astrocyte-secreted bFGF. Our findings indicate that long-term survival of human stem cell-derived cholinergic neurons requires trophic factors from nonneuronal cells. This data may provide insights into the development of an in vitro model of long-term cultured human cholinergic neurons useful for understanding of the mechanisms of cholinergic differentiation and developing treatments for neurological diseases. (c) 2007 Wiley-Liss, Inc.
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