Animal Husbandry and Procurement
Animal Health Surveillance
NSG Breeding Colony
Quarantine and Infectious Agent Containment
Training and Consultation
Transgenic Mouse Services
Veterinary Care and Pharmacy
Animal Facility (AF) staff coordinates animal procurement to ensure correct protocol assignment and tracking of animal numbers, and oversees routine husbandry procedures. Individually ventilated cages are changed bi-weekly under bio-safety cabinets. Routine monitoring of feed, water and animal health is conducted daily. Sick animals are reported and seen by consulting veterinarians.
A representative sampling of rodents is tested periodically for common murine viruses and mycoplasma using serological testing of a combination of breeder or experimental animals and sentinel animals. One mouse from 5% of the cages in each room is sent for serology assessment as well as in house pinworm and mite testing every six months. The results of these tests are filed and sent to the appropriate investigators. Sentinel rodents are housed on each cage rack side in every holding room and in the quarantine room to monitor the room health status. Every six months one sentinel from each cage is sent for serology, MPV PCR helicobacter, and endo-ecto parasitology. In addition, every six months one-two mice from our NSG colony are sent to Charles River Diagnostic testing for an HM basic, PCR for P. carinii, MPV PCR, helicobacter PCR, and endo-ecto parasites.
This service provides academically-oriented consultation time and scientific collaboration from an expert veterinary pathologist with expertise in mouse pathology and embryology. This service is free of charge to investigators at the The Wistar Institute Cancer Center. The goal is to foster and enhance scientific projects that involve animal models of cancer, inflammation, autoimmunity and transgenic and knockout technology, with the intention to publish innovative and academically sound research in premiere scientific literature.
Dr. David Garlick, DVM, board-certified veterinary pathologist with demonstrated expertise in mouse histopathology, will provide Wistar investigators with project consultation, histology slide evaluation and report preparation, and high-quality image development. See Dr. Garlick's areas of expertise.
Faculty who have pathology inquiries may email Dr. Garlick directly, providing a brief but detailed summary of their experimental protocol, key experimental questions being asked and material provided.
Upon receipt, Dr. Garlick will review the material, and provide a written pathology report along with high-quality images suitable for publication. In addition to email, which is the preferred method of communication, Dr. Garlick will be accessible for phone consultation with the originating investigators, if needed.
The AF staff maintains an NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) breeding colony that produces immunodeficient mice, lacking mature T cells, B cells, or functional NK cells, and which are deficient in cytokine signaling, allowing better engraftment of human cell types than any other strain. To place an order or request more information please contact us by phone (215) 898-3782/3780.
Only animals from approved vendors and that have been reviewed and approved by a veterinarian, are accepted into the barrier. Rodents that do not conform to these standards are placed in quarantine room or are excluded from the barrier if infected with any excluded agent. Quarantined animals are released only when they are proven free of any active murine virus infection, the helicobacter status is known, and pinworm and mite tests are negative. All mice in quarantine are treated as if they have pinworms and mites and are fed irradiated ivermectin/fenbendazole combination feed for eight weeks. A percentage of incoming mice are bled upon arrival and again at seven weeks, and are then sent to a diagnostic lab for serology testing of murine viruses. Helicobacter testing is also performed for each cage. Direct contact sentinels are housed in a proportion of the cages (determined by the veterinarian) for 28 days and are also sent to a diagnostic lab for serology, MPV PCR, and endo- and ecto-parasitology testing. Additional in house pinworm and mite testing is conducted at the end of the nine week quarantine holding period. If all the tests are confirmed negative at the end of the quarantine period, following review by a veterinarian, the rodents are released to a designated holding room.
The following are the steps and specific details for the quarantine process:
- Mice received are housed in quarantine and placed on an Ivermectin/Fendbendazole diet for 8 consecutive weeks.
- One mouse from each cage is ear tagged and bled upon arrival for the first bleed (mouse assessment). This provides our base line to confirm the murine status of the mice upon arrival.
- A fecal sample is taken from each cage for helicobacter testing.
- The mice from the initial bleed will be bled again for the 7-week bleed (mouse assessment). This bleed will show if any sero-conversion took place during transportation or while in quarantine.
- Ovariectomized CD-1 sentinels must be direct- housed with the mice for 28 days. When mice are received, any strains of the same sex with 5 mice are split so the sentinel can be with one cage and be within compliance of the cage space regulations for 28 days. Once the sentinels are sent to the diagnostic lab the mice are put back together into the original host cage. We will monitor any males very closely incase fighting occurs.
- The sentinels are sent to the diagnostic lab after they have been direct-housed with mice directly for 28 days and tested for serology - mouse assessment, MPV PCR, Direct exam for pinworm, mites, and lice and tape tested for endo-parasites.
- Breeding can begin in quarantine at 6-weeks. The lab notified to order the necessary cages so breeding cages can be set up. The new cages must be placed on the Ivermectin/Fendbendazole medicated diet.
- The mice are taken off the medicated diet at 8 weeks and placed on our standard diet.
- At 9-weeks, the mice are in-house tested for pinworm and mites (no cost to a PIs lab) by our animal health technicians.
- Testing Results are sent to PI and transfer procedures are arranged for the mice to enter the barrier with approval from the attending veterinarian.
AF staff is available to conduct special IACUC-approved procedures, including tumor measurements, ear tagging, and blood collection. Staff perform euthanasia upon request or when advised by veterinarians in the event of an animal welfare concern. Investigators are notified before euthanasia is performed and the AF staff will perform the euthanasia for investigators upon request. In addition, the staff performs animal health corrective action upon request when animal welfare concerns are reported. To request services or more information please contact us by phone, (215) 898-3782/3780.
Training is required and administered by the AF staff before new users may have independent access. The staff oversees training of users who are named on protocols, all of whom are required to attend an Animal Facility orientation, which provides instruction in societal concerns for the use of animals in research, existing laws, policies and guidelines regulating animal research, and current methods of animal care and use at Wistar. Users also complete an online training entitled “Working with the Wistar IACUC through the AALAS Learning Library”.
Professional Animal Health Technicians provide hands-on-training for all users to learn techniques for animal handling, sexing, blood collection, euthanasia and ensuring death. Injection training is available upon request. Specific IACUC mandated procedure training is available for users that require vet oversight and certificates. Veterinarians are also available for individual instruction on animal handling and experimental manipulations.
The Fox Chase Cancer Center (FCCC) transgenic mouse facility serves as the Transgenic Mouse resource for Wistar investigators. The aim is to provide access to technology and expertise for generating transgenic and genetically modified mice and to foster new scientific collaborations between researchers from both Institutions.
Benefits to Wistar researchers include, priority in scheduling, free consultations, and enhanced access to specialized facility services and to the expertise of Dr. Dietmer Kappes, Ph.D., Professor, FCCC Transgenic Mouse Facility Director, and Mr. Xiang (Sean) Hua, M.S., Manager.
How to Obtain Services
To obtain Transgenic Mouse services and pricing, please contact us by phone (215) 898-3782/3780 or Email: firstname.lastname@example.org. Access to transgenic and gene-targeting services is only available to Wistar investigators.
How It Works
Services are initiated when users contact the Wistar Animal Facility (AF). Facility staff then consults with the user to determine service requirements. Users may choose to have services performed either in the Wistar AF or in the transgenic mouse facility at FCCC. When services are provided at FCCC, the resulting products are delivered and enter into the WAF through our quarantine process. The following summarizes the workflow for transgenic mouse services:
1) Service provided within the Wistar AF:
User contacts the Wistar AF → Initial consultation → Wistar AF contacts the TMF at FCCC → Wistar AF, the User and the TMF coordinate → service provided and product delivered in the Wistar AF
2) Service provided within the TMF at FCCC:
User contacts the Wistar AF → Initial consultation → Wistar AF contacts the TMF at FCCC → Wistar AF, the User and the TMF coordinate → service begins in the TMF at FCCC → product delivered to the Wistar AF
Transgenic Mouse Facility Services and Technologies
Cryopreservation and revival of cryopreserved mouse lines
Cryopreservation of one line: Mouse lines cryopreserved as 8-cell stage embryos. Revival success rate from frozen embryos is 100%. Cryopreservation quality control is done by thawing and implanting the frozen embryos to witness the live birth, and this can be done for an additional fee at the PI's request (see 4303 below).
Revival of one cryopreserved mouse line: Frozen mouse embryos are thawed and implanted in recipient female mice that are generated on either FCCC or Wistar campus. A minimum of 5 live pups are guaranteed.
ES cell targeting
One ES cell microinjection session: One day of microinjection of ES cells into host blastocysts; typically inject 50-60 blastocysts on each day and most likely will obtain 3-5 chimeric males, depending on the ES clone quality. ES line 129 generally used. Will use C57BL6 ES cells on request. The efficiency of C57BL6 cells in chimera contribution as well as germ-line contribution is substantially lower than that of 129 cells.
ES cell targeting: Transfection, selection, and cryopreservation of 300-400 clones of “129” ES cells (W4 line from Taconic). Provide lysates of clones to lab for screening. Provide Transgenic Mouse Facility with clones from cryopreserved stocks designated by lab for blastocyst injection. Other mouse ES cells not currently utilized by the facility but may be upon request.
Transgenic mouse production
Pronuclear microinjection using DNA constructs into embryos (strain options include: B6C3F1, FVB/N, C57BL6) to produce transgenic mouse lines with a guarantee of certain number of founders based on the DNA construct size (e.g., 5 founders for DNA construct <10KB length, 3 for 10-20KB, 2 for 20KB+).
Single pronuclear microinjection: This option covers a single-day pronuclear microinjection work only, regardless of embryo strain, without any guarantee. Generally this is sufficient to obtain a sufficient number of founders. The facility should be consulted for more information.
Implant embryos into pseudopregnant recipient females generated from Wistar's own colony or from the colony at FCCC.
Technical service for minor procedures
Technical service: Consultations are provided without charge. Technical service covers all minor procedures in which the transgenic facility provides assistance.to PIs. Examples include micro-dissection, intrabursal injections, xenografting, micro surgeries, and more.
Two consulting veterinarians, Richard Boscoe, VMD, and Pierre Conti, VMD, MBA, DACLAM, are well-qualified and highly experienced to oversee veterinary care for the facility. One veterinarian (Dr. Boscoe) serves on the Wistar Cancer Center IACUC as the attending veterinarian, with the other (Dr. Conti) serving as alternate, assuring that the IACUC has veterinary expertise for review of each protocol. Each veterinarian is present for two-four hours at least one day per week (each Monday and each Thursday) to attend to any animal health issues that have been noted by the AF staff or the researchers. Vet reports are sent to the PI and lab for corrective action. At least one veterinarian is on call for emergencies at all times.
The AF staff manages a pharmacy providing agents for treatments prescribed by the veterinarians. The pharmacy also provides analgesics and anesthetics described in each protocol, with controlled substances distributed by designated staff under a veterinarian’s license.