CFP and YFP, but not GFP, provide stable fluorescent marking of rat hepatic adult stem cells.

Journal of Biomedicine & Biotechnology
Rouzbeh R Taghizadeh, James L Sherley

Abstract

The stable expression of reporter genes in adult stem cells (ASCs) has important applications in stem cell biology. The ability to integrate a noncytotoxic, fluorescent reporter gene into the genome of ASCs with the capability to track ASCs and their progeny is particularly desirable for transplantation studies. The use of fluorescent proteins has greatly aided the investigations of protein and cell function on short-time scales. In contrast, the obtainment of stably expressing cell strains with low variability in expression for studies on longer-time scales is often problematic. We show that this difficulty is partly due to the cytotoxicity of a commonly used reporter, green fluorescent protein (GFP). To avoid GFP-specific toxicity effects during attempts to stably mark a rat hepatic ASC strain and, therefore, obtain stable, long-term fluorescent ASCs, we evaluated cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP), in addition to GFP. Although we were unable to derive stable GFP-expressing strains, stable fluorescent clones (up to 140 doublings) expressing either CFP or YFP were established. When fluorescently marked ASCs were induced to produce differentiated progeny cells, stable fluorescence expression was...Continue Reading

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Citations

Nov 6, 2012·International Journal of Nanomedicine·Jin-Bao QinXin-Wu Lu
Jan 5, 2013·Sleep·Carlos Blanco-CenturionPriyattam J Shiromani
Dec 22, 2015·Applied Microbiology and Biotechnology·Samira SeifTahereh Taheri
May 15, 2015·Scientific Reports·Hugo R CairesCatarina R Almeida
Aug 29, 2013·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Jianhong WuYue Liu
Dec 31, 2011·Journal of Controlled Release : Official Journal of the Controlled Release Society·Jaesung KimSung Wan Kim
Oct 31, 2016·Hearing Research·J David MonroeAllison B Coffin
Jan 9, 2014·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Song HongQuansheng Wang
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Oct 28, 2019·Scientific Reports·Ken HashimotoM Charles Liberman
Jun 12, 2019·Bulletin of Experimental Biology and Medicine·S S VodopyanovA G Majouga
Sep 21, 2018·Scientific Reports·Benjamin E MaimonHugh M Herr

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Methods Mentioned

BETA
transgenic
transfection
density gradient centrifugation
flow cytometry
transfections
FACS

Software Mentioned

OpenLab
CellQuest
Summit

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