Establishment of a novel method for the production of chimeric mouse embryos using water-in-oil droplets.
Production of chimeric animals is often a necessity for the generation of genetically modified animals and has gained popularity in recent years in regenerative medicine for the reconstruction of xenogeneic organs. Aggregation and injection methods are generally used to produce chimeric mice. In the aggregation method, the chimeras are produced by co-culturing embryos and stem cells, and keeping them physically adhered, although it may not be an assured method for producing chimeric embryos. In the injection method, the chimeras are produced by injecting stem cells into the zona pellucida using microcapillaries; however, this technique requires a high degree of skill. This study aimed to establish a novel method for producing chimeric embryos via water-in-oil droplets that differs from conventional methods. In this study, embryonic stem cells and embryos were successfully isolated in the droplets, and the emergence of chimeric embryos was confirmed by co-culture for 6 h. Using this method, the control and operability of stem cell numbers could be regulated, and reproducibility and quantification were improved during the production of chimeric embryos. In addition to the conventional methods for producing chimeric embryos, the novel method described here could be employed for the efficient production of chimeric animals.
['Imai|Hiroyuki|H|', 'Tsuda|Soichiro|S|', 'Iwamori|Tokuko|T|', 'Kano|Kiyoshi|K|', 'Kusakabe|Ken Takeshi|KT|', 'Ono|Etsuro|E|']
D000818:Animals D030801:Animals, Genetically Modified D002478:Cells, Cultured D002678:Chimera D018920:Coculture Techniques D046149:Embryo Culture Techniques D004622:Embryo, Mammalian D053595:Embryonic Stem Cells D005260:Female D051379:Mice D009821:Oils D033581:Stem Cell Transplantation D014867:Water D015044:Zona Pellucida
embryos droplet chimera embryonic stem cell