RRC ID 176
Author Mothe AJ, Kulbatski I, van Bendegem RL, Lee L, Kobayashi E, Keating A, Tator CH.
Title Analysis of green fluorescent protein expression in transgenic rats for tracking transplanted neural stem/progenitor cells.
Journal J. Histochem. Cytochem.
Abstract Green fluorescent protein (GFP) expression was evaluated in tissues of different transgenic rodents--Sprague-Dawley (SD) rat strain [SD-Tg(GFP)Bal], W rat strain [Wistar-TgN(CAG-GFP)184ys], and M mouse strain [Tg(GFPU)5Nagy/J]--by direct fluorescence of native GFP expression and by immunohistochemistry. The constitutively expressing GFP transgenic strains showed tissue-specific differences in GFP expression, and GFP immunohistochemistry amplified the fluorescent signal. The fluorescence of stem/progenitor cells cultured as neurospheres from the ependymal region of the adult spinal cord from the GFP SD and W rat strains was assessed in vitro. After transplantation of the cells into wild-type spinal cord, the ability to track the grafted cells was evaluated in vivo. Cultured stem/progenitor cells from the SD strain required GFP immunostaining to be visualized. Likewise, after transplantation of SD cells into the spinal cord, immunohistochemical amplification of the GFP signal was required for detection. In contrast, GFP expression of stem/progenitor cells generated from the W strain was readily detected by direct fluorescence both in vitro and in vivo without the need for immunohistochemical amplification. The cultured stem/progenitor cells transplanted into the spinal cord survived for at least 49 days after transplantation, and continued to express GFP, demonstrating stable expression of the GFP transgene in vivo.
Volume 53(10)
Pages 1215-26
Published 2005-10
DOI 10.1369/jhc.5A6639.2005
PII jhc.5A6639.2005
PMID 15983120
MeSH Animals Animals, Genetically Modified Cells, Cultured Female Fluorescence Green Fluorescent Proteins / biosynthesis* Green Fluorescent Proteins / genetics Immunohistochemistry Male Mice Rats Rats, Sprague-Dawley Rats, Wistar Spinal Cord / cytology* Stem Cell Transplantation* Stem Cells / metabolism*
IF 2.37
Times Cited 33
Rats W-Tg(CAG-GFP)184Ys(strainID=525)