Cryo Service Frequently Asked Questions (FAQs)

What is the CMMR?
What is FIMRe?
What can I deposit to the CMMR?
How do I deposit my mutants and samples in the CMMR?
What do you mean by proven breeders?
How do I prepare my males for sperm collection?
How many breeders should I have in my colony before starting cryopreservation?
What can I withdraw from the CMMR?
What mutants are available at the CMMR?
How can I access mutants and samples for other repositories?
How do I withdraw mutants and samples from the CMMR?
Who can withdraw samples?
What conditions may apply?
What is an MTA?
What is the timeline for withdrawal and shipment of my samples?
How should your mutants and/or samples be shipped to the CMMR?
How much will it cost?
How do you calculate your charges?

What is the CMMR?
The Canadian Mouse Mutant Repository, CMMR, is a facility dedicated to the preservation and distribution of mutant mouse lines, samples, and ES cells (see CMMR-NorCOMM FAQs).

What is FIMRe?
The Federation of International Mouse Repositories (FIMRe) is a collaborating group of Mouse Repository and Resource Centres worldwide whose collective goal is to archive and provide strains of mice as cryopreserved embryos and gametes, ES cell lines, and live breeding stock to the research community. The CMMR is an active member of FIMRe.

What can I deposit to the CMMR?
Mice: Investigators can deposit mutant mouse lines in the form of embryos, gametes, or ES cell lines. Cryobiology specialists at the CMMR receive, process, cryopreserve, and archive the mutants and samples. Services rendered and service fees depend on the type and number of samples cryopreserved. An investigator may also deposit previously frozen mouse embryos, gametes or ES cell lines and pay applicable processing and storage fees. The CMMR can also perform mutant mouse strain expansion and/or recovery for experimental purposes.

Tissues & Cells: An investigator can deposit whole fixed tissues or cells and fixed tissues or cells on glass slides for storage, subject to applicable storage fees. The CMMR, in conjunction with the adjacent CMHD Pathology Core, can receive live mice from investigators and recover and fix and process tissues and cells for subsequent storage at our repository in conjunction with mouse strain cryopreservation services.

How do I deposit my mutants and samples in the CMMR?
The Depositing Investigator (DI) should start the process by registering for an account in the TCP LIMS and setting up a Mouseline passport for the strain(s) to be deposited. These passports provide information about the derivation, specific mutation(s), genetic background, genotyping protocol(s), coat colour, phenotype and health status of the strain. Guidelines naming transgenic and mutant strains can be found in Rü licke et al. (2007) Laboratory Animals41:301-311. Mouseline passport information will be checked for accuracy and completeness.

Once a Mouseline Passport has been completed, the DI should complete the appropriate service request form in the TCP LIMS. Our instruction page details the steps involved in completing a request. Once a service request is submitted, a Material Transfer Agreement (MTA) provided by the CMMR is executed between the CMMR at the Hospital for Sick Children and the DI or his/her institution to guarantee ownership of his/her samples. Once the MTA is complete, the service will be scheduled. The CMMR will receive, process, cryopreserve, and archive the mutants or samples per our fee schedule. The DI will have the option of making his/her mutants or samples available to other investigators.

Go to instruction page

What do you mean by proven breeders?
A proven breeder male has fathered at least two litters of pups, in which the gene(s) of interest has been detected. The two litters may be from one or two females.

A proven breeder female has given birth to at least one litter of pups, in which the gene(s) of interest have been detected.

How do I prepare my males for sperm collection?
The highest quality sperm is harvested from young males, 8-16 weeks old, that have been housed apart from females for at least three days.

How many breeders should I have in my colony before starting cryopreservation?
For Speed Cryo, you should have at least three productive breeding cages in your colony prior to starting. You can start Speed Cryo by shipping two proven breeder males to the CMMR. Once those males have been used to generate embryos, you can ship another breeder male to complete the process. Males used for sperm collection are not recoverable for subsequent breeding.

For all other cryopreservation services, be sure that you maintain at least one productive breeding cage in your colony until cryopreservation is complete.

What can I withdraw from the CMMR?
Mice: An investigator can withdraw cryopreserved mutant mice in the form of embryos, gametes or ES cells. The CMMR can also provide an investigator with live mice derived from any of the above cryopreserved specimens. Service charges depend on the form of mutant mice provided to the investigator. The CMMR can also perform strain expansion and recovery for experimental purposes.

ES Cells: Please see CMMR-NorCOMM FAQs.

Tissues & Cells: An investigator can request whole fixed tissues or cells and/or fixed tissues or cells on glass slides, subject to applicable processing fees.

What mutants are available at the CMMR?
Mice: Investigators can search our catalogue of ENU and non-ENU mutants. Detailed information about gene mutations, alleles, or genetic background can be found through MGI.

ES Cells: Investigators can search the NorCOMM production facility web accessible databases through the Centre for Modeling Human Disease (CMHD) or the Mammalian Functional Genomics Centre (MFGC) websites.

Tissues & Cells: Coming soon.

How can I access mutants and samples for other repositories?
The CMMR is an active member of the Federation of International Mouse Repositories (FIMRe). Investigators are able to request samples from FIMRe members. Samples are shipped from the originating repository to the CMMR for recovery as live mice and distribution to the Withdrawing Investigator/Institution. Shipping of gametes and cells across international borders is often less restricted than shipping live mice. The CMMR has considerable experience in the recovery of mutant mice from all cryopreserved forms. Details on how to recover mutant strains from FIMRe members can be found on our Services page. Please contact the CMMR for further details.

How do I withdraw mutants and samples from the CMMR?
The Withdrawing Investigator or Institution (WI) initiates the process by registering for an account in the TCP LIMS. Once registered, the WI completes a Service Request form to recover or withdraw their mutant of interest as outlined on our instruction page . Depending on the sample(s) requested, a Material Transfer Agreement (MTA) provided by the CMMR may have to be executed prior to processing the request. The MTA describes the conditions associated with the use of mutants and samples and confirms the rights of the Depositing Investigator/Institution (DI). For mutants and samples from FIMRe members, the MTA will be provided by the originating repository. In some cases, not-for-profit DIs will be asked to accept the Conditions of Use rather than execute a MTA.

Requests for frozen samples will be fulfilled by dispatching cryopreserved mutants and samples in appropriate shipping containers. Frozen samples are normally shipped on dry ice or in dry shippers via overnight courier at the WI’s expense. Dry shippers must be returned to the CMMR by the WI as soon as possible to enable their repeated use. Requests for live animals require recovery from cryopreserved samples (if applicable) and co-ordination of animal shipments between the CMMR and the WI’s animal facility.

Go to instructions page

Who can withdraw samples?
Most recipients will be associated with a recognized biomedical or genetics research institution. Depositing Investigators and/or their institutions may restrict the distribution of their strains and samples.

What conditions may apply?

The Depositing Investigator or Institution (DI) retains ownership of the sample(s). The CMMR acts with the DI to distribute sample(s) and notifies the DI before releasing samples from the archive. The DI may prohibit access to an archived sample. All DIs will receive updated inventories of their mutants and samples at the CMMR, along with a list of all Withdrawing Investigators/Institutions ( WIs) who have received their mutants and/or samples. The DI will be responsible for the authenticity of the mutants and samples and any relevant documentation.

It is the responsibility of the WI to ensure that any executed MTAs enable their research goals and to obtain appropriate licenses for any relevant patents or other uses. Further it is the responsibility of the WI to ensure that the WI signatory(ies) on any MTA has(have) the legal authority to sign the MTA.

International WIs must conform to their country’s rules and regulations for importing or exporting biological samples and/or live animals. It is the WI’s responsibility to inform the CMMR of any regulations pertaining to or special documentation required for the shipment of requested samples or animals.

All WIs must agree to conform to the relevant governing regulations concerning the use of biological reagents and the ethical treatment of animals.

What is an MTA?
A Material Transfer Agreement (MTA) is an agreement between two or more parties for the transfer of mutants and/or samples. MTAs generally cover issues pertaining to the ownership, restrictions of use, intellectual property and commercialization of the exchanged mutants and/or samples. MTAs are legally binding documents and must be signed by either the investigator or his/her institution, whosoever has the legal authority to sign such documents. The CMMR uses MTAs as the formal agreement for transferring mouse strains from Depositing Investigators/Institutions (DIs) to the CMMR and from the CMMR to Withdrawing Investigators/Institutions ( WIs). MTAs must be fully executed prior to sample deposition or withdrawal.

What is the timeline for withdrawal and shipment of my samples?

Mice: After receiving a request for withdrawal and the completion of all paperwork associated with a request, the CMMR will contact the Withdrawing Investigator/Institution (WI) to set up a schedule for reanimation of mice from cryopreserved strains or for the shipment of cryopreserved strains/samples from the CMMR archive to the WI. The timelines below are from the commencement of reanimation using cryopreserved germ cells or embryos stored at the CMMR. Additional time may be required to obtain suitable recipient mice, suitable germ cell donor mice from commercial suppliers, fresh sperm or live donor mice from investigators, or when repeated attempts are necessary because of low reanimation efficiency. The scheduled start of reanimation attempts will depend on the number of requests in the queue.

Each request for withdrawal will be given a specific date of commencement so that the WI will have an idea of the length of time required to obtain mice. CMMR staff will keep the WI informed of the experimental progress.

Recovery of Live Mice…

…from Frozen Embryos : Aapproximately 6-7 weeks for weaned pups to be produced and ready for shipping.

…from Frozen Ovaries: Approximately 10-11 weeks after ovary transplant, the first litter of pups may be produced and ready for shipping. Alternatively, the transplant recipients can be shipped to the WI for breeding and maintenance at their animal facility.

…from Fresh/Frozen Sperm and ICSI: Weaned pups may be available for shipping as early as 8 weeks after commencing reanimation. However, it is common for ICSI to require multiple attempts at embryo production and this could extend the timeline for production of live mice, particularly with C57BL/6 sperm.

…from Fresh/Frozen Sperm and Unassisted IVF: Approximately 8 weeks for weaned pups to be produced and ready for shipping.

Somatic tissues and cells: For tissues and/or cells in stored in the CMMR archive, it will take a few days for the items to be shipped out to the Withdrawing Investigator.

How should the mutants and samples be shipped to the CMMR?

Live Animals: Contact us for details

Somatic tissues or Cells: Each sample should be placed in individually labeled containers. The samples should be packaged and prepared according to sample type (contact us for details).

How much will it cost?
Please refer to the CMMR Service Charges.

How do you calculate your charges?
Base fees are calculated on a cost recovery basis. Fees charged to Investigators at the Hospital for Sick Children are subsidized by overhead paid by the Hospital for Sick Children on behalf of the CMMR. Therefore, charges to SickKids investigators are somewhat less than those charged to other requesters.