Generation of ES cell chimeras
The Investigator provides two vials of frozen ES cells suitable for seeding on 1 well of a standard 6-well cluster plate. If the ES cells have not been generated by the Core, documentation about the absence of mycoplasm contamination, karyotype information and culture conditions must be provided by the Investigator. The Core will prepare the ES cells for injection into C57Bl/6J blastocysts and will inject at least 50 blasts per injection experiment. Offspring will be turned over to the Investigator at weaning age (3-4 weeks). Successful production of chimeras is only guaranteed if the ES cells have been generated by the Core. If no offspring are obtained, the experiment is repeated at no cost to the Investigator.
To initiate an experiment, the Investigator contacts the TG Core Supervisor and submits a completed order form for blastocysts injections. Injections will be performed at the earliest possible time, usually beginning within 4 weeks after receipt of the ES cells. Offspring will be born 3 weeks after injection and tail snips may be obtained 3 weeks thereafter for screening. Fee information is available from the TG Core Supervisor.
Other procedures can on special occasions be arranged in collaboration with the Investigator: this includes embryo aggregation experiments, embryo-ES cell aggregation experiments, and generation of tetraploid embryo-ES cell aggregation chimeras.