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EMBRYONIC STEM CELLS

Efficient Production of Mice from Embryonic Stem Cells Injected into Four- or Eight-Cell Embryos by Piezo Micromanipulation

^aCollege of Life Sciences, Sun Yat-Sen University, Guangzhou, China;
^bDepartment of Obstetrics and Gynecology, University of South Florida College of Medicine, Tampa, Florida, USA;
^cCollege of Life Science, Nankai University, Tianjin, China

Key Words. Embryonic stem cells • Piezo microinjection • Mice • Pluripotency

Correspondence: David L. Keefe, M.D., Department of Obstetrics and Gynecology, University of South Florida College of Medicine, 12901 Bruce B. Downs Boulevard, Tampa, Florida 33612, USA; e-mail: dkeefe{at}health.usf.edu; or Lin Liu, Ph.D., University of South Florida College of Medicine, 12901 Bruce B. Downs Boulevard, Tampa, Florida 33612, USA; e-mail: liutelom{at}yahoo.com

Received February 19, 2008; accepted for publication April 28, 2008.
First published online in STEM CELLS EXPRESS   May 8, 2008.

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.

Disclosure of potential conflicts of interest is found at the end of this article.