Analysis of green fluorescent protein expression in transgenic rats for tracking transplanted neural stem/progenitor cells.
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.
Division of Cellular and Molecular Biology, Toronto Western Research Institute, Ontario, Canada
Andrea J
AJ
Mothe
Iris
I
Kulbatski
Rita L
RL
van Bendegem
Linda
L
Lee
Eiji
E
Kobayashi
Armand
A
Keating
Charles H
CH
Tator
D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D005260:Female; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D013116:Spinal Cord / Q000166:cytology*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000378:metabolism*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins