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)
Sperm freezing protocol CARD gCPA (Sep 2018)
Sperm freezing protocol from epididymides following refrigerated transport (Sep 2018)
IVF protocol using sperm from epididymides following refrigerated transport (Sep 2018)
IVF protocol using freshly harvested and frozen sperm (MBCD+GSH) (Sep 2018)
IVF protocol for sperm samples frozen in vials and MTG straws(Sep 2018)
Laser-Assisted IVF (Sep 2018)
Mouse intracytoplasmic sperm injection (ICSI) (Nov 2016)
A simple and efficient vitrification procedure for cryopreservation of mouse embryos in straws (Sep 2018)
Harvest and cryopreservation of in vivo derived mouse embryos (Sep 2018)
Thawing and use of cryopreserved mouse embryos (Sep 2018)
Non- Surgical Embryo Transfer (Sep 2018)
Acid Tyrode’s Treatment of Oocytes (Sep 2018)
Preparation of Media and Reagents (Sep 2018)
Consumables & Supplies (Sep 2018)
Selected References (Sep 2018)
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.