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Embryo Services

Service Details

Service Charges

How To Order


Service Details

Embryo Cryopreservation

Embryo Cryopreservation archives embryos for long-term storage. This service is used to protect against genetic drift, preserve mouselines not currently needed as live mice, enable distribution of a mouseline, and to serve as a back-up against mouseline loss due to a reduction in breeding productivity or loss of a mouseline. Depending on the specific Embryo Cryopreservation service selected, this process will preserve specific gene mutations and/or genetic background. We recommend that 200-400 embryos, depending on embryo genotype, be cryopreserved to ensure recovery of the line and enable distribution, if applicable. For all cryopreservation services Quality Control to Live Born is essential to ensure that the mouseline is recoverable before the strain is discontinued or distributed.

It is important to note that oocyte, embryo, and sperm harvest are terminal procedures. By special request, collection of other tissues for experimental purposes can be arranged through the CMHD Pathology Core, located next door to the CMMR at the TCP. For a description of CMHD Pathology Services and Fees please seehttp://www.phenogenomics.ca/services/pathology.html?v=3.

Basic Cryo

Embryos are produced by matings overseen by the Requester in their own animal colony. Requesters set up timed matings between stud males and superovulated females in their colony. Superovulation protocols are age-dependent and for most strains effective in very young females (3 weeks for most strains). Hence, it is often more time- and cost-effective to breed mutant males to superovulated wild-type femalesof appropriate strain background to maximize embryo numbers. If strain background and/or homozygosity are critical, female mice from the investigator's colony may be used. We recommend that at least 8-10 proven breeder males be available as studs for each Basic Cryo request. Depending on available number of males, their fertility and strain response to superovulation, up to four sessions of Basic Cryo may be necessary to produce adequate embryo numbers for effective mouseline archiving.

Requesters provide up to 10 mated 2.5-dpc pregnant females to the CMMR for embryo harvest and cryopreservation. Suggestions, breeding protocols and gonadotropin hormones for superovulation to assist the Requester can be provided if necessary. If the Requester's colony is at TCP, TCP staff can oversee superovulation, mating and transfer of plugged females for embryo harvest and cryopreservation. Additional fees apply for these services and are outlined below.

Each session of Basic Cryo includes embryo harvest and cryopreservation from up to 10 plugged females in one session. The number of embryos produced depends on the fertility of the strain and whether superovulation protocols exist and/or are used before mating. Embryos are stored in straws, with ~20 embryos per straw, in liquid nitrogen. Depending on mating scheme, homozygous, heterozygous, or both genotypes of embryos are preserved. Both genetic background and mutations of interest are preserved if males and females are from the mouseline of interest.

Choose Basic Cryo when you have good fertility and plug rates in your colony and have at lease 8 stud males to dedicate to mating for embryo harvest. While females will be transferred for embryo harvest, males may be maintained in the Requester's colony, provided females are available there or wild-type females can be imported to the colony as required.

Service fees include embryo harvest and cryopreservation. The number of cryopreservation sessions needed depends on the number, genotype, and quality of embryos collected.

Service fees do not include transport of mice from the Requester to the CMMR, animal housing at the TCP, if required, or embryo storage fees, if applicable. For each strain cryopreserved by Basic Cryo, at least one Quality Control to Live Born session is necessary to ensure that the mouseline is recoverable before the mouseline is discontinued or distributed. Additional charges apply for quality control to live born.

Speed Cryo

In vitro fertilization (IVF) is used to produce large numbers of embryos for cryopreservation. Requesters provide up to two males, eight to 35 weeks old, per IVF session as sperm donors for fertilization of wild-type oocytes. Best sperm counts are obtained from relatively young proven breeder males that have been separated from females for a few days before scheduled sperm harvest.

Each Speed Cryo request includes one IVF session, using sperm harvested (see sperm cryopreservation for details of sperm harvest) from one or two males to fertilize oocytes isolated from 20 superovulated wild-type females. Two-cell stage embryos are frozen after 24 hours of in vitro culture. Ten to twenty embryos are cultured for an additional 48-72 hours to determine in vitro developmental potential. Oocyte donors are purchased from commercial vendors and the following wild-type strains may be selected by the Requester: C57BL/6J, C57BL/6NCrl, C57BL6/NTac,C3H, 129, CD-1, FVB or ICR. Embryos are frozen in straws, with ~20 embryos/straw. The actual number of embryos produced depends on the fertility of the males provided and the strain of female oocyte donors. For best results, proven male breeders should be provided for Speed Cryo. Depending on genotype of the male(s), either heterozygous or a mix of heterozygous and wild-type embryos will be preserved.

For each mouseline cryopreserved by Speed Cryo at least one session of Quality Control to Live Born is necessary to ensure that the mouseline is recoverable before the mouseline is discontinued or distributed. Additional charges apply for quality control to live born. Also for an additional charge, some sperm can be cryopreserved in parallel, if requested at the same time as Speed Cryo.

Choose Speed Cryo when you want to dedicate a small number of mice from your colony for cryopreservation, you do or can maintain your mutation(s) of interest on a wild-type background, and you can accept one generation of mating after embryo cryorecovery to produce homozygous mutant mice.

Service fees include one IVF session, embryo harvest, and cryopreservation, using donor male sperm and wild-type oocytes, with in vitro developmental assessment of 10-20 embryos. The number of IVF sessions needed depends on the number, genotype and quality of embryos collected.

Service fees do not include the purchase and receipt of oocyte donors, animal housing charges for either female or male donors, quality control to live born, or embryo storage fees, if applicable.

Embryo Quality Control to Live Born

Quality control protocols are in place for all embryo cryopreservation procedures. For each mouseline cryopreserved, one straw of embryos is thawed and embryo viability is assessed in vitro. Viable embryos are then transferred to one or two pseudopregnant females and housed until pups are born. This will provide an indication of the number of embryos required to ensure recovery of the line.

Service fees include recovery of previously frozen embryos and transfer into pseudopregnant recipient.

Service fees do not include animal housing or tissue sampling of pups for genotyping.

Embryo Storage

Embryo batches are divided and stored at two different sites in storage vessels under 24-hr surveillance to ensure safety of the frozen archive.

Embryo Cryorecovery

Embryo Cryorecovery is used to reanimate mouselines previously cryopreserved as embryos. This procedure can be done with embryos from CMMR stock or from other repositories or facilities. Transport of embryos is often less expensive that shipping live animals and relieves the animal welfare concerns associated with shipping live animals. Embryo Cryorecovery derives animals into SPF status at TCP.

From CMMR Stock

The Principal Investigator (PI) (and designate) must have an account on TCP LIMS and complete the following steps to request Embryo Cryorecovery from CMMR stock:

Obtain approval of a new Animal Use Protocol (AUP) or amendment(s) to an existing AUP for the mouseline(s) being recovered so that the recovered mice will be accepted at the TCP or home animal facility for housing. If mice or embryos are being shipped across international borders, the PI must ensure that all paperwork applicable to animal or embryo export/import is in order prior to the mice or embryos being shipped.

The PI or designate must complete a Service Request form on TCP LIMS entitled Cryo Recovery from Embryos before the CMMR will schedule cryorecovery.All CMMR stocks have existing TCP LIMS Mouseline Passports to enable service request entry.
Once scheduled, embryos will be thawed and transferred to pseudopregnant females.

Service fees are per straw of embryos and include the recovery of embryos from CMMR stock, thawing the embryos, and one embryo transfer surgery to a single pseudopregnant recipient. If necessary due to embryo number, additional pseudopregnant recipients may be needed for an additional charge.

Service fees do not include clipping or genotyping of pups, individual health testing of foster mother, if required, animal housing, or shipping costs.

From FIMRe or Other Repositories

Cryopreserved embryos from other repositories can also be recovered at the CMMR. This is helpful when mouselines are located outside of Toronto as shipping and import of embryos is often easier than that of live mice. For all requests to the CMMR, the PI and designate, if applicable, must have a registered account on TCP LIMS. Several steps must be completed by the Principal Investigator (PI) or designate:

Complete and have approved all paperwork associated with the importation of embryos into Canada, including any Material Transfer Agreements, CMMR Embryo Import Form(s), customs, and animal import forms required. The PI should forward copies of all forms required to accompany the shipment to the originating repository.

The PI or designate should place and pay for the order of embryos. Once all paperwork and financial items have been arranged, the PI or designate should put the supplying repository in touch DIRECTLY with CMMR staff (lauryl.nutter@phenogenomics.ca) so that the frozen embryos from the supplying repository are shipped directly to the CMMR. All shipping is at the PI's expense unless included in the costs paid to the supplying repository. In any case, a courier account number should be provided to the CMMR for any shipping expenses not covered. These include the cost of return shipping of any dry shippers used to transport frozen embryos.

The PI or designate completes a Service Request form on TCP LIMS entitled Cryo Recovery from Embryos before the CMMR will schedule shipping of the embryos. For mouselines new to the TCP LIMS, this will require that the PI or designate complete a Mouseline Passport for the mouseline prior to completing the Service Request form. Please see Rülicke et al. (2007) Laboratory Animals 41:301-311 for guidelines for nomenclature of genetically-engineered mice.

After embryos arrive, the service will be scheduled and the embryos thawed and transferred to pseudopregnant recipient(s) into SPF health status at the TCP.

Service fees are per straw of embryos and include communications between the CMMR and supplying repository, the receipt of embryos from the supplying repository, thawing the embryos, and a single embryo transfer into a pseudopregnant recipient. If necessary due to embryo number, additional pseudopregnant recipients may be needed for an additional charge.

Service fees do not include charges from supplying repository, clipping and genotyping of pups, health testing of foster mother, animal housing, or shipping costs.

Embryo Straw Withdrawal

Shipping of cryopreserved germ cells and embryos, especially across international boundaries, is often simpler than shipping live mice. As a result, investigators who have the local ability to recover live mice from cryopreserved embryos or germ cells can purchase cryopreserved stock from our publicly available mouselines.

Service fees are per straw and include the withdrawal and packaging of cryopreserved mouselines from CMMR stock.

Service fees do not include export administration fee, if applicable, or shipping, which is done via the courier and account number provided by the investigator at the time of their request.

Service Charges

Service SickKids TCP Investigators Non-Profit Canadian Institution Non-Profit International Institution
Embryo Cryopreservation
Basic Cryo, per cryo session, TCP F1 or F2

$370

$481

$481

$592

Containment or External

$432

$562

$562

$691

Speed Cryo, per IVF session TCP F1 or F2

$730

$915

$950

$1170

Containment or External

$805

$1015

$1045

$1290

QC to Live Born, per attempt
  • Includes recovery of previously frozen embryos and transfer into pseudopregnant recipient
  • Can be requested in conjunction with any embryo cryopreservation service
  • Does not include animal housing, tissue collection or genotyping

$280

$305

$361

$445

Additional Embryo Transfer, per recipient
  • Includes embryo transfer surgery for each pseudopregnant recipient
  • Can occur in conjunction with QC to Live Born or any cryorecovery service
  • Does not include animal housing, health testing of individual recipients, tissue collection or genoytping

$95

$95

$124

$152

Superovulation and mating for Basic Cryo, up to 10 mice
  • Includes hormones, injection, mating and plug checking of one up to 10 females
  • Does not include animal costs
TCP F1

$115

$115

$150

$184

TCP F2

$165

$165

$215

$264

Hormones, per 5IU aliquot
  • Includes PMS and hCG hormones needed for standard dosing of 5 mice

$8

$8

$10

$13

Annual Storage Charge, per mouseline
  • Storage of up to 20 straws of embryos at two different sites in monitored liquid nitrogen storage tanks

$30

$39

$39

$48

Embryo Cryorecovery
Embryo Cryorecovery, per straw from CMMR
  • Includes recovery of frozen embryos from CMMR stock and transfer to one pseudopregnant recipient
  • Does not include material cost of embryo straw, if applicable, clipping or genotyping pups, health testing of individual recipients, animal housing or shipping

$470

$585

$610

$750

Embryo Cryorecovery, per straw from other repository
  • Includes communication with supplying repository, receipt and temporary storage of embryos, embryo recovery, and transfer to one pseudopregnant recipient
  • Does not include fees charged by supplying repository, shipping, clipping or genotyping pups, health testing of individual recipients, or animal housing

$605

$760

$785

$965

Additional Embryo Transfer, per recipient
  • Includes embryo transfer surgery for each pseudopregnant recipient
  • Can occur in conjunction with QC to Live Born or any cryorecovery service
  • Does not include animal housing, health testing of individual recipients, tissue collection or genoytping

$95

$95

$124

$152

Embryo Straw, per straw from CMMR stock
  • Cost of embryo straw from available CMMR stock
  • Does not include sample export administration fee, if applicable

$220

$220

$286

$320

Use of Dry Shipper , per use
  • Includes use of dry shipper, labels, and arranging shipping for transport of embryos or sperm
  • Does not include liquid nitrogen to charge dry shipper or shipping costs

$75

$97.50

$97.50

$120

Charging Dry Shipper, each charge
  • Includes liquid nitrogen and labour to charge dry shipper for transport of embryos or sperm
  • Does not include use of dry shipper or shipping costs

$30

$39

$39

$48


How to Order

Please follow the Order Instructions with the relevant Order Form below.

Service Service Request Form
Embryo Cryopreservation
Basic Cryo Basic Embryo Cryopreservation
Speed Cryo Speed Cryopreservation of Embryos
Embryo Cryo Recovery
From CMMR Stock Cryo Recovery from Embryos
From FIMRe or Other Repositories Cryo Recovery from Embryos
Embryo Straw Germ Cell and Embryo Withdrawal

 

 

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