Resusciation

Resuscitation of mouse lines uses frozen germplasm (embryos or sperm, IVF). This is the preferred method to choose when importing lines from outside OHSU. Given a choice, we recommend sperm to be imported as the chance of success is much higher.

In Vitro Fertilization (IVF)

We will order 15 WT female mice of the required strain for superovulation and fertilization using the designated frozen sperm sample. If possible, the PI could provide two tubes (straws) of frozen sperm, with an additional tube (straw) as a backup.

We will transfer fertilized embryos generated by IVF into up to four CD1 recipient females.

Resuscitation via Frozen Embryo Transfer

Our team will thaw one aliquot of frozen embryos transfer them into one to two CD1 recipient females. The quality of the frozen embryos provided by the PI is a critical factor for success. Therefore, successful resuscitation cannot be guaranteed.

Requesting resuscitation services

To request a resuscitation project please go to the AMSC iLab page and fill in the necessary information there.

There will be an additional form that we will provide to reserve space with Department of Comparative Medicine (DCM) in their specific-pathogen free (SPF) quarantine room before we begin your project. 

1. We ask that you request two samples of germplasm if possible.
2. Once you have arranged an MTA with the sending partner, please place us in contact with them to make shipping arrangements. We have a dry shipper available for rental to help expedite the process.
3. Once we receive the samples, we will transfer them into our LN2 dewer for storage until we can schedule your procedure.

Rederivation

Rederivation is used to remove pathogens from a strain so that it can move from a conventional mouse room to a specific-pathogen free (SPF) room. It can take one of two forms:

1. In vitro fertilization using sperm harvested from males of the desired genotype to fertilize embryos from twenty wildtype donors. This will result in embryos transferred into 3-4 foster dams.
2. Standard rederivation where breeding age males are paired with super-ovulated females age 3-4 weeks either provided by the PI or ordered by the core depending on the desired results. The embryos are harvested and transferred into a number of foster dams depending on the number of embryos retrieved. Typically, this is only 1-2 dams but does result in homozygous offspring in the case of females provided by the PI. Different strains of mice have varying reactions to super ovulation so there is no guarantee of success.

Requesting rederivation services

To request a rederivation project please go to the AMSC iLab and fill in the necessary information there.

There will be an additional form that we will provide to reserve space with the Division of Comparative Medicine (DCM) in their SPF quarantine room before we begin your project.

In addition, you will need to provide the following animals:

1. 4-6 stud males from the line you are rederiving.
2. 5-6, 3-4 week old mutant females from the line you are rederiving if you are going the in vivo route (if you want to have homozygous mouse stock, otherwise we can order WT  females)

On occasion we may perform rederivations from imported live animals. Please note that this is subject to approval by DCM based on animal health reports, and approval for the project must first be obtained via a DCM import request form.

Standard rederivation service fees include

• Purchase of 3 week old females (donors of oocytes).
• Superovulation of 3 week old females.
• Collection and Transplantation of rederived embryos into pseudopregnant foster females.
• The production of guaranteed clean animals.
• Animal housing charges for fosters and their offspring until weaning.

Service fees do not include:

• Ordering of special strains of female mice for oocyte donors (see additional services below).
• Animal housing charges for litters after weaning.
• Ear/tail tissue collection for DNA isolation for genotyping.
• Health testing of the foster mother and pups.
• Shipping charges.

Additonal services

At your request AMSC can purchase and use other inbred mouse lines for the rederivation of mutant mice. Strains that cost significantly more than C57BL/6J will be charged an additional fee based on the difference in cost.