Quantification of green fluorescent protein by in vivo imaging, PCR, and flow cytometry: comparison of transgenic strains and relevance for fetal cell microchimerism.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
Yutaka FujikiKirby L Johnson

Abstract

Animal models are increasingly being used for the assessment of fetal cell microchimerism in maternal tissue. We wished to determine the optimal transgenic mouse strain and analytic technique to facilitate the detection of rare transgenic microchimeric fetal cells amongst a large number of maternal wild-type cells. We evaluated two strains of mice transgenic for the enhanced green fluorescent protein (EGFP): a commercially available, commonly used strain (C57BL/6-Tg(ACTB-EGFP)10sb/J) (CAG) and a newly created strain (ROSA26-EGFP) using three different techniques: in vivo and ex vivo fluorescent imaging (for whole body and dissected organs, respectively), PCR amplification of gfp, and flow cytometry (FCM). By fluorescent imaging, organs from CAG mice were 10-fold brighter than organs from ROSA26-EGFP mice (P < 0.0001). By PCR, more transgene from CAG mice was detected compared to ROSA26-EGFP mice (P = 0.04). By FCM, ROSA26-EGFP cell fluorescence was more uniform than CAG cells. A greater proportion of cells from ROSA26-EGFP organs were positive for EGFP than cells from CAG organs, but CAG mice had a greater proportion of cells with the brightest fluorescent intensity. Each transgenic strain possesses characteristics that make it...Continue Reading

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Citations

Sep 29, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Anna J Studwell, Darrell N Kotton
Jul 29, 2010·Molecular Human Reproduction·Eddy S M LeeNicholas M Fisk
Jun 8, 2012·Biology of Reproduction·Stephanie PritchardDiana W Bianchi
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Feb 26, 2013·Journal of Reproductive Immunology·E SeppanenK Khosrotehrani
Aug 30, 2008·Neuropharmacology·Elena V RomanovaJonathan V Sweedler
Nov 15, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Attila Tárnok
Nov 6, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Laura TeodoriAttila Tárnok
Jul 8, 2011·Journal of Reproductive Immunology·Lisa R KallenbachKirby L Johnson
Jun 25, 2017·Proceedings of the National Academy of Sciences of the United States of America·Rodney M RitzelLouise D McCullough
Oct 14, 2017·Briefings in Functional Genomics·Anders StåhlbergThomas Kroneis
Jan 26, 2018·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Dema NajemMahmud Bani-Yaghoub
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Feb 24, 2018·Scientific Reports·Raquel CastosaAngélica Figueroa
Mar 27, 2015·Nano Letters·Ryan M WilliamsDaniel A Heller
Apr 14, 2020·Organic Letters·Rachael FayJason P Holland

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