Monitoring of Venus transgenic cell migration during pregnancy in non-transgenic rabbits.
Cell transfer between mother and fetus were demonstrated previously in several species which possess haemochorial placenta (e.g. in humans, mice, rats, etc.). Here we report the assessment of fetal and maternal microchimerism in non-transgenic (non-TG) New Zealand white rabbits which were pregnant with transgenic (TG) fetuses and in non-TG newborns of TG does. The TG construct, including the Venus fluorophore cDNA driven by a ubiquitous cytomegalovirus enhancer, chicken ß-actin promoter (CAGGS), was previously integrated into the rabbit genome by Sleeping Beauty transposon system. Three different methods [fluorescence microscopy, flow cytometry and quantitative polymerase chain reaction (QPCR)] were employed to search for TG cells and gene products in blood and other tissues of non-TG rabbits. Venus positive peripheral blood mononuclear cells (PBMCs) were not detected in the blood of non-TG littermates or non-TG does by flow cytometry. Tissue samples (liver, kidney, skeletal and heart muscle) also proved to be Venus negative examined with fluorescence microscopy, while histology sections and PBMCs of TG rabbits showed robust Venus protein expression. In case of genomic DNA (gDNA) sourced from tissue samples of non-TG rabbits, CAGGS promoter-specific fragments could not be amplified by QPCR. Our data showed the lack of detectable cell transfer between TG and non-TG rabbits during gestation.
NARIC-Agricultural Biotechnology Institute, Gödöllő, Hungary. liptak.nandor@abc.naik.hu.
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Lipták
NARIC-Agricultural Biotechnology Institute, Gödöllő, Hungary.
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Hoffmann
NARIC-Agricultural Biotechnology Institute, Gödöllő, Hungary.
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Kerekes
NARIC-Agricultural Biotechnology Institute, Gödöllő, Hungary.
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Iski
Department of Pharmaceutical Biotechnology, University of Pécs, Pécs, Hungary.
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Ernszt
Department of Pharmaceutical Biotechnology, University of Pécs, Pécs, Hungary.
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Kvell
NARIC-Agricultural Biotechnology Institute, Gödöllő, Hungary.
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Hiripi
NARIC-Agricultural Biotechnology Institute, Gödöllő, Hungary.
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Bősze
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000254:growth & development; D002465:Cell Movement / Q000235:genetics*; D002645:Chickens / Q000235:genetics; D046528:Chimerism / Q000196:embryology*; D004251:DNA Transposable Elements / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D033261:Maternal-Fetal Relations; D008856:Microscopy, Fluorescence; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000235:genetics; D011817:Rabbits
D004251:DNA Transposable Elements
CAGGS promoter; Microchimerism; New Zealand white rabbits; Non-transgenic; Sleeping Beauty transposition; Venus protein