Cryopreservation protocols
All EMMA partners use validated protocols for mouse embryo and spermatozoa freezing techniques, in vitro fertilisation procedures and embryo transfers. However, the EMMA partners use different protocols that are optimized for their individual settings. EMMA distribution centres will provide to customers ordering frozen embryos or spermatozoa the centre specific validated thawing protocols. The protocols provided for download below are practiced in EMMA training courses at the MRC-MGU (Harwell) and are routinely used by the Harwell EMMA node.
The Harwell node further provides a number of training videos that enable members of the scientific community to learn the skills and techniques that are used at the MRC. A series of short films that take you step-by-step through the process of freezing mouse embryos and sperm for use in IVF can be viewed at the MRC-Harwell website.
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Introductory presentation on cryopreservation (Feb 2013)
Mouse sperm cryopreservation and IVF recovery - MBCD+GSH protocol (Feb 2013)
Mouse IVF recovery using freshly harvested sperm - MBCD+GSH protocol (Feb 2013)
Rescue IVF protocol for frozen sperm (Feb 2013)
Mouse sperm cryopreservation from epididymides after transportation at refrigerated temperatures (Jan 2015)
Laser-assisted IVF protocol (Feb 2013)
Mouse intracytoplasmic sperm injection (ICSI) (Nov 2016)
Mouse embryo cryopreservation (Feb 2013)
Vitrification of mouse embryos (May 2016)
Vitrification of mouse embryos in a big batch (Jul 2016)
Thawing and use of cryopreserved embryos (Feb 2013)
Transcervical embryo transfer (TCET) (Feb 2016)
Acid tyrodes treatment of oocytes (Aug 2009)
Cryopreservation media and reagents (Feb 2013)
Cryopreservation consumables and supplies (Feb 2013)
EMMA partners have also developed and implemented innovative shipment protocols that facilitate fast and flexible exchange processes, reduce costs and provide alternatives to live animal transportation. Current SOPs for shipping refrigerated spermatozoa in Lifor to archiving centres, and SOPs for shipping refrigerated embryos and shipping frozen sperm on dry ice are listed below.
The protocol on the production of germ-free mice is provided by the Gulbenkian Institute which is offering the EMMA axenic (germ-free) service.