Efficient production of mice from embryonic stem cells injected into four- or eight-cell embryos by piezo micromanipulation.
The conventional method for producing embryonic stem (ES) cell-derived knockout or transgenic mice involves injection of ES cells into normal, diploid blastocysts followed by several rounds of breeding of resultant chimeras and thus is a time-consuming and inefficient procedure. F0 ES cell pups can also be derived directly from tetraploid embryo complementation, which requires fusion of two-cell embryos. Recently, F0 ES cell pups have been produced by injection of ES cells into eight-cell embryos using a laser-assisted micromanipulation system. We report a simple method for producing F0 ES cell germline-competent mice by piezo injection of ES cells into four- or eight-cell embryos. The efficiency of producing live, transgenic mice by this method is higher than that with the tetraploid blastocyst complementation method. This efficient and economical technique for directly producing F0 ES cell offspring can be applicable in many laboratories for creating genetically manipulated mice using ES cell technology and also for stringent testing of the developmental potency of new ES cell or other types of pluripotent stem cell lines.
Huang
Junjiu
J
Department of Obstetrics and Gynecology, University of South Florida College of Medicine, 12901 Bruce B. Downs Boulevard, Tampa, Florida 33612, USA.
Deng
Kai
K
Wu
Haojia
H
Liu
Zhong
Z
Chen
Zhisheng
Z
Cao
Shanbo
S
Zhou
Lingjun
L
Ye
Xiaoying
X
Keefe
David L
DL
Liu
Lin
L
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002470:Cell Survival; D046149:Embryo Culture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D008297:Male; D051379:Mice; D008846:Micromanipulation / Q000379:methods*; D018895:Microsatellite Repeats; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic; D013997:Time Factors; D019076:Transgenes