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

2. An International Centre for Mouse Genetics Mary Lyon Centre – high barrier unit

3. An International Centre for Mouse Genetics 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. An International Centre for Mouse Genetics CNR/IBC Istituto di Biologia Cellulare, Monterotondo, Italy CNRS/CDTA Centre de Distribution, de Typage et d'Archivage animal, Orléans, France MRC/MGU Mammalian Genetics Unit, Harwell, UK Karolinska Institutet Karolinska Institutet, Stockholm, Sweden FCG/IGC Instituto Gulbenkian de Ciência, Oeiras, Portugal HMGU/IEG Institute of Experimental Genetics, Munich, Germany EMBL/EBI European Bioinformatics Institute, Hinxton, UK GIE-CERBM/ICS Institut Clinique de la Souris, Illkirch/Strasbourg, France Sanger Institute Wellcome Trust Sanger Institute, Hinxton, UK CNB/CSIC Centro 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 European Mouse Mutant Archive - EMMA

5. An International Centre for Mouse Genetics An international service

6. An International Centre for Mouse Genetics Host bi-annual cryocourses  Hands on demonstration of:  Embryo freezing  Sperm freezing  In vitro fertilization  Reference point  Disseminate skills

7. An International Centre for Mouse Genetics 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. An International Centre for Mouse Genetics Transport - Dry shipper  Keep samples at LN 2 Temp  Expensive  Re-usable  Considered safe by IATA  Robust

9. An International Centre for Mouse Genetics What we cryopreserve  Pre-implantation embryos  Oocytes  Spermatozoa  Ovarian tissue

10. An International Centre for Mouse Genetics Cryopreservation of the pre- implantation embryo  Controlled rate freezing  Vitrification

11. An International Centre for Mouse Genetics Freezing machines Planer Kryo 10 FTS - BioCool IV

12. An International Centre for Mouse Genetics Goblet Clamp CBS goblets - nice idea but!

13. An International Centre for Mouse Genetics Sperm freezing equipment

14. An International Centre for Mouse Genetics Cooling Rate Measuring cooling rates Comark thermocouple used to measure freezing rates Temperature during transportation not measured a present (Thermoguard or iButtons)

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

16. An International Centre for Mouse Genetics Mean±SD (n=5) Cooling Rate LN 2 h Cooling rates – dewar

17. An International Centre for Mouse Genetics GroupsCooling rate (°C/min)MotileProgressiveRapid BOX-37.4027.2510.5014.25 35CM-7023.258.2513.50 30CM-115.8846.7512.2519.75 25CM-145.8855.2514.7523.00 Cooling R Cooling rates – effects on fertility