Linked Life Data

18205194

Source:http://linkedlifedata.com/resource/pubmed/id/18205194

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2008-2-4
pubmed:abstractText
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 useful under specific experimental circumstances. The CAG mouse model is preferable when experiments require brighter cells, whereas ROSA26-EGFP is more appropriate when uniform or ubiquitous expression is more important than brightness. Investigators must carefully select the transgenic strain most suited to the experimental design to obtain the most consistent and reproducible data. In vivo imaging allows for phenotypic evaluation of whole animals and intact organs; however, we did not evaluate its utility for the detection of rare, fetal microchimeric cells in the maternal organs. Finally, while PCR amplification of a paternally inherited transgene does allow for the quantitative determination of rare microchimeric cells, FCM allows for both quantitative and qualitative evaluations of fetal cells at very high sensitivity in a plethora of maternal organs.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-11778067, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-12095299, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-12781134, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-15949558, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-17074776, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-17269129, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-17598185, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-17721542, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-17912040, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-18307256, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-325766, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-7268890, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-8570620, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-9175875, http://linkedlifedata.com/resource/pubmed/commentcorrection/18205194-9461201
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
1552-4930
pubmed:author
pubmed:copyrightInfo
(c) 2008 International Society for Analytical Cytology
pubmed:issnType
Electronic
pubmed:volume
73
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
11-118
pubmed:dateRevised
2010-12-3
pubmed:meshHeading
pubmed:year
2008
pubmed:articleTitle
Quantification of green fluorescent protein by in vivo imaging, PCR, and flow cytometry: comparison of transgenic strains and relevance for fetal cell microchimerism.
pubmed:affiliation
Division of Genetics, Department of Pediatrics, Tufts-New England Medical Center, Boston, Massachusetts, USA.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural