The University of Queensland
Brisbane, Queensland 4072
Australia

Email:e.williams@imb.uq.edu.au
Ph +61 7 33462276
Fx +61 7 33462101

©TASQ 2005
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The production of Knock Out mice by aggregation of embryonic stem cells with morula stage mouse embryos can be routinely done here in TASQ.

The zona pellucida is removed from the embryos by washing through several drops of Tyrods Acid solution. The ES-cells are prepared and allowed to re-aggregate to form clumps. One clump of ES-cells is added to each embryo. The following day the embryos should have developed to blastocyst stage. These embryos are transferred, resulting in chimeric pups.

The aggregation technique can also be used for making embryo-embryo chimeras. These can be mutant, wild-type, normal diploid or tetraploid embryos.

Prices:

  • $AU 900 / session (internal)
  • $AU 1200 / session (external)

This price covers:

  • Superovulated and mated CD1 donor females
  • Embryo collection
  • Morula aggregation
  • Embryo culture
  • Pseudopregnant mothers and embryo transfer
  • Animal agisment charges for pseudopregnant mothers and pups till weaning

Additional Notes and Recommendations

  • TASQ recommends 2 intitial sessions for each clone.
  • TASQ will ask investigators to provide 2 frozen vials of clones for injection. These clones should have had a chromosome count and only those proven with a 70% of a 40 count will be accepted. TASQ will prepare cells for injection.
  • A TASQ Application Form for Morula Aggregation must be completed prior to the commencement of service.
  • Additional agistment costs apply once pups are weaned and for the transport of mice if applicable.
  • AEC and GTR numbers must be supplied prior to commencement of service.
  • TASQ will test chimaera for germline transmission at an additional cost to client.
  • Penalty fees apply to late cancellations of Aggregation sessions.
  • The service being provided is undertaken with no guarantee of the genetically manipulated mouse strain.

Preparation of ES Cells

Morula Aggregation References

Khillan, J.S. and Bao, Y. (1997) Preparation of animals with a high degree of chimerism by one-step coculture of embryonic stem cells and preimplantation embryos. Biotechniques. 22, pp 544-549.

Kong, W. et al (2000) Preparing transgenic animals with a simplified method of morula aggregation using ES cells. Lab Animal. 29, 25-30.