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An overview of the MRC-Harwell’s cryopreservation service

An overview of the MRC-Harwell’s cryopreservation service. Martin Fray Frozen Embryo & Sperm Archive – (FESA) Medical Research Council Harwell, UK. Mary Lyon Centre – high barrier unit. The MRC frozen embryo archive. Worldwide Genetic Resource of mouse models of human disease

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An overview of the MRC-Harwell’s cryopreservation service

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  1. An overview of the MRC-Harwell’s cryopreservation service Martin Fray Frozen Embryo & Sperm Archive – (FESA) Medical Research Council Harwell, UK

  2. Mary Lyon Centre – high barrier unit

  3. The MRC frozen embryo archive • Worldwide Genetic Resource of mouse models of human disease • ~1450 stocks, >470,000 embryos • Includes transgenic, mutants, chromosome anomalies & inbred strains • Sole UK archiving centre • http://www.har.mrc.ac.uk • EMMA (European Mouse Mutant Archive)

  4. European Mouse Mutant Archive - EMMA CNR/IBCIstituto di Biologia Cellulare, Monterotondo, ItalyCNRS/CDTACentre de Distribution, de Typage et d'Archivage animal, Orléans, France MRC/MGUMammalian Genetics Unit, Harwell, UKKarolinska InstitutetKarolinska Institutet, Stockholm, Sweden FCG/IGCInstituto Gulbenkian de Ciência , Oeiras, PortugalHMGU/IEGInstitute of Experimental Genetics, Munich, GermanyEMBL/EBIEuropean Bioinformatics Institute, Hinxton, UK GIE-CERBM/ICSInstitut Clinique de la Souris, Illkirch/Strasbourg, France Sanger InstituteWellcome Trust Sanger Institute, Hinxton, UK CNB/CSICCentro Nacional de Biotecnología, Madrid, Spain Fleming Biomedical Sciences Research Centre Al. Fleming, Athens, Greece OULU University of Oulu, Oulu, Finland BIAT Vetmeduni Vienna, Biomodels Austria, Vienna, Austria IMG Institute of Molecular Genetics, Prague, Czech Republic

  5. An international service

  6. Host bi-annual cryocourses • Hands on demonstration of: • Embryo freezing • Sperm freezing • In vitro fertilization • Reference point • Disseminate skills

  7. Liquid nitrogen storage • Samples retained in mobile dewars (n=35) • Bulk (Chart) LN2 tanks (n=10) • 3 x LN2 rooms all with an independent LN2 supply • BOC and Cryo-service deliver to site

  8. Transport - Dry shipper • Keep samples at LN2 Temp • Expensive • Re-usable • Considered safe byIATA • Robust

  9. What we cryopreserve • Pre-implantation embryos • Oocytes • Spermatozoa • Ovarian tissue

  10. Cryopreservation of the pre-implantation embryo • Controlled rate freezing • Vitrification

  11. Freezing machines • Planer Kryo 10 • FTS - BioCool IV

  12. Goblet Clamp CBS goblets - nice idea but!

  13. Sperm freezing equipment

  14. Cooling Rate Measuring cooling rates • Comark thermocouple used to measure freezing rates • Temperature during transportation not measured a present (Thermoguard or iButtons)

  15. Cooling Rate Cooling rates – Thermosafe box Sperm straw in container Styrofoam 2.5 cm 6-9 cm Liquid nitrogen Mean±SD (n=3)

  16. Cooling Rate Cooling rates – dewar h LN2 Mean±SD (n=5)

  17. Cooling R Cooling rates – effects on fertility

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