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Characterization of Human Neural Progenitor Cell Models for Developmental Neurotoxicity Screening
Citation:
SHAFER, T. J., J. M. BREIER, AND W. R. MUNDY. Characterization of Human Neural Progenitor Cell Models for Developmental Neurotoxicity Screening. Presented at DNT 2 Meeting, Reston, VA, November 12 - 14, 2008.
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Description:
Current testing methods for developmental neurotoxicity (DNT) make evaluation of the effects of large numbers of chemicals impractical and prohibitively expensive. As such, we are evaluating two different human neural progenitor cell (hNPC) models for their utility in screens for DNT. ReNcell CX (ReN CX) cells are a c-myc immortalized hNPC line that differentiates into neurons and glia in the absence of growth factors. In the presence of EGF and FGF, these cells expressed the neuroprogenitor markers nestin and SOX2, confirming their neuroprogenitor cell nature. Cell proliferation (BrdU incorporation) assays were developed in cells treated with a selected set of antiproliferative compounds and effects were compared to effects on viability (propidium iodide staining); each endpoint was quantified using an automated, high-content imaging approach. When effects of diverse set of 16 chemicals was examined on proliferation and viability using this approach, 6 of 8 compounds known to be developmentally neurotoxic decreased proliferation and/or viability, while only 1 of 8 presumably “non-toxic” (based on a lack of reported effects in the peer-reviewed literature) compounds altered these endpoints. ENstem-A cells are a neuroprogenitor cell model derived from the NIH-approved H9 human embryonic stem cell line that has not been exogenously immortalized. These cells express the NPC markers nestin, and SOX-2, with low levels of the oligodendrocyte marker Oct-4. We recently have obtained this cell line and expanded this cell line and are now beginning to develop assays for proliferation and cytotoxicity. By comparison to ReNcell CX cells, this cell line is not immortalized by a myc transduction. Overall, the goal of this work is to determine the most appropriate human cell model(s) for use in high throughput screens for developmental neurotoxicity. (This is an abstract of a proposed presentation and does not necessarily reflect EPA policy).