Cellutions 2009 Volume 2
Characterization of MilliTrace CX nestin GFP Reporter Human neural stem Cell Line
Christine Chen, Ph.d. and Vi Chu, Ph.d., Millipore Corporation, Temecula, Ca
Nestin, an intermediate filament protein, is a well-known marker that is expressed in proliferating neural progenitors and stem cells during the early stages of development in the CNS and PNS. The regulatory element of the Nestin gene is widely used as a promoter to drive expression of GFP in transgenic mice. These transgenic mice models provide an opportunity to study neural stem cell (NSC) development in vivo and in vitro. A human GFP NSC reporter cell model would be a valuable tool to examine cellular function during neurogenesis. Toward this end, a human cortical neural stem reporter cell line was established in which the expression of humanized mulleri GFP (hmGFP) was placed under the regulation of the mouse Nestin promoter. We show that the mouse Nestin promoter has been stably integrated into the cell genome and that the resulting Nestin GFP reporter human neural stem cells faithfully recapitulated the expression profile of Nestin in vitro, namely, GFP fluorescence remained constant during NSC propagation and steadily declined upon differentiation. The down regulation of GFP fluorescence makes it possible to monitor neuronal differentiation in live cells.
Keywords: Stem Cells