Simple and Efficient Production of Embryonic Stem Cell-Embryo Chimeras by Coculture

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 10; pp. 4582 - 4585
• Main Authors: Wood, Stephen A., Pascoe, Wendy S., Schmidt, Christel, Kemler, Rolf, Evans, Martin J., Allen, Nick D.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.05.1993; National Acad Sciences; National Academy of Sciences
• Subjects: Animals; Biological and medical sciences; Biology; Blastocyst; Blastocyst - cytology; Cell lines; Cells, Cultured; Chimera; Chimeras; Chimerism; Coculture techniques; Embryo, Mammalian - cytology; Embryology: invertebrates and vertebrates. Teratology; Embryonic stem cells; Embryos; Feeder cells; Female; Fetuses; Fundamental and applied biological sciences. Psychology; General aspects; General aspects. Development. Fetal membranes; Genetics; Germ cells; Germ Cells - cytology; In Vitro Techniques; Male; Mice; Mice, Inbred Strains; Rodents; Stem cells; Embryonic development; Embryo; Stem cell; Rodentia; Blastocyst; Method; Vertebrata; Mixed cell culture; Mammalia; Mouse; Germ line; Chimerism; Chimera
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.10.4582

A method for the production of embryonic stem (ES) cell-embryo chimeras was developed that involves the simple coculture of eight-cell embryos on a lawn of ES cells. After coculture, the embryos with ES cells attached are transferred to normal embryo culture medium and allowed to develop to the blastocyst stage before reimplantation into foster mothers. Although the ES cells initially attach to the outside of the embryos, they primarily colonize the inner cell mass and its derivatives. This method results in the efficient production of chimeras with high levels of chimerism including the germ line. As embryos are handled en masse and manipulative steps are minimal, this method should greatly reduce the time and effort required to produce chimeric mice.