eagle-i The University of PennsylvaniaThe University of Pennsylvania
See it in Search
This page is a preview of the following resource. Continue onto eagle-i search using the button on the right to see the full record.

Mouse Line Rederivation

eagle-i ID

http://eagle-i.itmat.upenn.edu/i/0000016d-d5f4-7a8e-eaa8-15d780000000

Resource Type

  1. Material production service

Properties

  1. Fee for service
    Yes
  2. Resource Description
    Mouse line rederivation is used in many institutions as a method of re-establishing you mouse line or as a means to eliminate pathogenic or potentially pathogenic agents. This procedure involves transferring pre-implantation stage zygotes into recipient surrogate mothers (for completion of development). Fertilized zygotes for rederivation can be generated in several different ways, described below: –Harvest and freeze embryos; This option is performed for complex (more than 2 mutant alleles) mouse lines housed in ARC, that do not possess the required health status to enter the CTRB animal facility. Your males (minimum of 5 mice, 7 weeks old+) are mated with your females (5 mice, 3-5 weeks old), the resulting fertilized zygotes are harvested and cryopreserved. A day or two later the zygotes are thawed and transferred into surrogate mother mice housed in CTRB. When the rederived offspring reach 10-14 days old they will be tail snipped, the tissues will be delivered to your lab and screened for the appropriate genotype. Mice identified with the correct genotype will be transferred to your animal holding room for future breeding and experimental use. -Pre-frozen embryos; The fastest and most efficient method for importing a mouse line is to have pre-frozen embryos shipped directly to the Transgenic Core. We will receive the shipment, store appropriately until needed and regenerate the mouse line. Alternatively, the strain may already be archived in our storage units. To recover the strain, vials are removed from liquid nitrogen, the zygotes are slowly thawed and rehydrated then the zygotes are transferred into surrogate mother mice housed in CTRB. When the rederived offspring reach 10-14 days old they will be tail snipped, the tissues will be delivered to your lab and screened for the appropriate genotype. Mice identified with the correct genotype will be transferred to your animal holding room for future breeding and experimental use. -Cooled sperm; This service is ideal for importing strains from institutions who do not have access to cryopreservation facilities. Reproductive tissues from 1-2 male mice are shipped to the Transgenic Core at 4 degrees. Upon receipt of the samples the Core will perform In-Vitro Fertilization (IVF) with oocytes isolated from wild-type donors. Fertilized zygotes are transferred into surrogate mother mice housed in CTRB. When the rederived offspring reach 10-14 days old they will be tail snipped, the tissues will be delivered to your lab and screened for the appropriate genotype. Mice identified with the correct genotype will be transferred to your animal holding room for future breeding and experimental use. This service requires a strict schedule, due to the viability of the sperm, and the essential timing involved when performing IVF. -Pre-frozen sperm; The pre-frozen sperm can be imported directly to the Transgenic Core. We will receive the shipment, store appropriately until needed and regenerate the mouse-line. Alternatively, the strain may already be archived in our storage units. To recover the strain, vials are removed from liquid nitrogen, the sperm is slowly thawed, allowed to recover then used for IVF with oocytes, isolated from wild-type donors. The following day viable zygotes are transferred into surrogate mother mice housed in CTRB. When the rederived offspring reach 10-14 days old they will be tail snipped, the tissues will be delivered to your lab and screened for the appropriate genotype. Mice identified with the correct genotype will be transferred to your animal holding room for future breeding and experimental use. -Freeze and thaw sperm for IVF; This option is performed for less complex mouse lines housed in ARC, that do not possess the required health status to enter the CTRB animal facility. Tissues are removed from 1-3 males and the sperm is isolated and frozen. Approximately a week later a sample of the sperm is thawed and used for IVF with oocytes isolated from wild-type donors. Fertilized zygotes are transferred into surrogate mother mice housed in CTRB. When the rederived offspring reach 10-14 days old they will be tail snipped, the tissues will be delivered to your lab and screened for the appropriate genotype. Mice identified with the correct genotype will be transferred to your animal holding room for future breeding and experimental use. The method of rederivation you choose to adopt should be determined based on the genotype of the animals and your goals with the strain. Strains involving 2 or more allele modifications ideally should not be rederived using in-vitro fertilization options. The zygote transfer methods (A or B) would produce mice with the correct genotype much faster than the sperm options. All strains of mice can be rederived, but the efficiency of the procedure depends on the hardiness and the fertility rate of the strain. Inbred strains may occasionally require the use of more mice and could take exceptionally more time. After the zygotes have been surgically transferred into the reproductive tracts of the surrogate mother mice they will remain in the care of the transgenic core. The surrogates will deliver the pups 3 weeks post-transfer. Approximately 2 weeks later the Core will isolate tail snips for your lab to genotype. The offspring will be weaned when they are 3 weeks old. At this time the Veterinary Department may decide to perform some health assessments on the animals to verify their health status. This procedure will take approximately one to two weeks to complete. After which, the animals can be transferred to your animal holding room.
  3. Service Fee URL
    https://corelabs.research.chop.edu/sites/default/files/inline-files/Transgenic_Fee_Schedule_9.25.19.pdf
  4. Service Provided by
    Transgenic Core (CHOP)
  5. Website(s)
    https://corelabs.research.chop.edu/transgenic/services
 
RDFRDF
 
Provenance Metadata About This Resource Record
  1. workflow state
    Published
  2. contributor
    ggrant (Gregory Grant)
  3. created
    2019-10-16T15:06:40.166-04:00
  4. creator
    ggrant (Gregory Grant)
  5. modified
    2019-10-16T15:07:33.006-04:00
Copyright © 2016 by the President and Fellows of Harvard College
The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016