Simple and efficient production of mice derived from embryonic stem cells aggregated with tetraploid embryos.
Six newly derived hybrid mouse embryonic stem (ES) cell lines and two inbred ES cell lines were tested for their ability to produce completely ES cell-derived mice by aggregation of ES cells with tetraploid embryos. Forty-five ES cell-tetraploid pups were generated from six hybrid ES cell lines and no pups from two inbred ES cell lines. These pups were found to have increased embryonic and placental weights than control mice. Twenty-two pups survived to adulthood and produced normal offsprings, and the other 23 pups died of several reasons including respiratory distress, abdomen ulcer-like symptoms, and foster failure. The 22 adult ES cell-tetraploid mice were completely ES cell-derived as judged by coat color and germline transmission, only two of them was found to have tetraploid component in liver, blood, and lung as analyzed by microsatellite loci. Our data suggested that genetic heterozygosity is a crucial factor for postnatal survival of ES cell-tetraploid mice, and tetraploid embryo aggregation using hybrid ES cells is a simple and efficient procedure for immediate generation of targeted mouse mutants from genetically modified ES cell clones, in contrast to the standard protocol, which involves the production of chimeras and several breeding steps.
Li
Xiangyun
X
College of Life Science, Peking University, Beijing, China. lixiangyun35@yahoo.com.cn
Yu
Yuansong
Y
Wei
Wei
W
Yong
Jun
J
Yang
Jie
J
You
Jiefang
J
Xiong
Xiaoran
X
Qing
Tingting
T
Deng
Hongkui
H
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002459:Cell Fusion; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D051379:Mice; D011003:Ploidies; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*