Division of Laboratory Animal Resources (DLAR)

UK Core Rederivation, Cryopreservation, and Barrier Facility

Facility Description

The University of Kentucky Sanders Brown Embryo Cryopreservation, Rederivation Core (SBECRC) and Barrier Facility is part of the Division of Laboratory Animal Resources (DLAR). The facility consists of approximately 5,000 net ft2 facility located on the fourth floor of the Sanders-Brown Center on Aging. The facility was renovated through funding by the National Institutes of Health and became operational in 2013.

The Sanders Brown barrier facility is managed by Jeanie Kincer, DVM, DACLAM and the core facility services are primarily performed by Sharon Giles, a veterinary technician with 24 years of experience in the field. Sharon has attended the Jackson Laboratory Workshops on Mouse Colony Management, Embryo Transfer, and Cryopreservation of Mouse Germplasm and the use of the NSET device for non-surgical embryo transfer. She has also attended the Taconic Germ Free Seminar and Workshop on the use and maintenance of both germ-free isolators and axenic and gnotobiotic mouse colonies. Animal husbandry personnel (DLAR) provide additional support as needed.

Services Offered

  • Rederivation of contaminated mouse strains via embryo transfer. 

Many strains of mice carry clinical or subclinical infectious agents that may become major pathogens in immunodeficient mice.  Additionally, a number of these pathogens may exert a subtle impact on specific research studies. The UK-SBECRC has the capability to eliminate these pathogens from colonies of mice via in-house embryo transfer.

  • High-level barrier housing capabilities to permit maintenance of nuclear colonies, breeding, and expansion of rederived colonies.

Following embryo rederivation and successful elimination of pathogens it is critical that the newly rederived strain be housed and handled so as to prevent reintroduction of the pathogens. Returning the initial small number of the animals to the investigator for expansion greatly increases the risk of inadvertent contamination since the investigator and research staff are typically still working with infected animals on research projects. The UK-SBECRC high-level barrier facility permits colony breeding and expansion under conditions minimizing the potential of re-infection. Once the colony reaches an adequate size, the investigator can make a rapid transition to using only the clean animals, greatly reducing the potential for inadvertent contamination and greatly reducing research efforts delays.

  • Embryo and sperm cryopreservation capabilities along with storage options for maintaining special and unique mouse strains in long-term storage. 

Embryo and sperm cryopreservation are well established techniques in mice used to maintain strains without continual breeding and to provide a backup in the advent of a catastrophic event such as natural disaster, breeding cessation or disease outbreak. The in-house cryopreservation capabilities offered by the UK-SBECRC allow UK investigators to preserve valuable strains and provide an emergency backup source of the strains should a catastrophic loss of the breeding colony occur. Cryopreservation is also more cost-efficient than maintenance of strains through live animal breeding, allowing researchers to conserve precious research dollars.

  • Gnotobiotic and Axenic isolator capabilities.

The role of the microbiota in disease pathogenesis and host response is an area of increasing research interest. The ability to house animals in gnotobiotic or axenic conditions requires very specialized equipment, extensive sterilization capabilities, and expertise. This new facility will provide the capability to house mice in defined microbial conditions. Four positive pressure semi-rigid isolators will provide an isolated environment in which the animals may be housed and evaluated. These isolators are intended primarily to be study-specific with a high level of investigator involvement and use. The UK-SBECRC will provide the support services (sterilization) and assistance needed to maintain the isolators.

  • Per Diem Rates:

For questions regarding rates and fees please contact facility personnel using the information below.

Contact Information

Anyone with any further questions or with any interest in a tour of the facility and its services should contact either Sharon or Dr. Kincer.

  • Sharon Giles, Veterinary Technician
    Phone: 859-218-1439
    Email: sharon.giles3@uky.edu

  • Jeanie Kincer, DVM, DACLAM
    Phone: 859-323-5469
    Email: jeanie.kincer@uky.edu

  • Facility/Contact Hours:
    Monday - Friday, 7am - 4pm

Extended Facility Description

  • Functional Design Description

The facility essentially consists of three separate units, supported by a central support core, and functioning as an integrated facility.  The embryo rederivation and cryopreservation facility (yellow) consists of a quarantine room, a necropsy-embryo collection station, a laboratory area for embryo manipulation and evaluation, controlled rate freezers and liquid nitrogen storage for embryo cryopreservation, and a pass-through BSC to permit transfer of the embryos into the barrier embryo implantation laboratory for rederivation.

The high-level barrier facility (green) consists of 3 barrier animal housing rooms. The first and smallest barrier room will house the embryo transfer recipient dams and litters. Following weaning of the litter and confirmation of microbial status, the weaned pups will be transferred to one of two barrier housing rooms. Each room has the capability of housing up to 500 cages of mice. All equipment and supplies entering the barrier housing rooms will be sterilized and showers and sterile clothing will be required of all individuals entering the barrier housing facility. 

The gnotobiotic isolator room (blue) is the final functional component and consists of a gowning room for entrance and a single large isolator housing room. All supplies and equipment to the isolators will be sterilized in the central support facility and delivered to the gnotobiotic isolator room. 

Major equipment items in the Central Support Core (reds and browns) include a rack washer, bottle filler, bedding dispenser, large pass-through chlorine dioxide sterilization chamber and 2 large pass through floor-loading steam autoclaves. Areas for cage dumping and washing along with clothes washing facilities, and feed and bedding storage are also provided off the central corridor.

Sanders Brown Personnel Flow

  • Traffic Flow

The facility has strictly controlled traffic flow pattern. Personnel enter into the main support corridor through a security card access-controlled door. Research personnel using the gnotobiotic isolator facility will access the facility from the stairway and directly enter the gowning room across the hall (blue arrows). After changing into the appropriate protective gowns they will enter via another controlled access door into the gnotobiotic isolator facility. Personnel egress from the facility is the reverse of entrance. Equipment flow is completely separate from personnel flow in the gnotobiotic isolator facility.

Personnel entering the high level animal housing barrier and the sterile-support area of the facility will access the area through vestibule and restroom entrance (brown arrow). Card access is required to access the vestibule and to exit the vestibule to the sterile corridor area. All personnel entering through the vestibule will be required to shower in the adjacent restrooms and change into sterile work attire prior to entering the sterile corridor (red arrow). Once in the sterile corridor personnel can enter any animal housing room and return to the sterile corridor (red arrows). Movement from the animal housing rooms to the return corridor is directional and re-entry is not permitted without showering and changing work attire (dark red arrows). Sterile support equipment and supplies for the gnotobiotic isolator facility can also be provided from the sterile corridor through the unidirectional support door connecting the two areas.

Personnel accessing the embryo cryopreservation and rederivation portion of the facility may enter directly from the main support corridor (yellow arrows). Personnel entering the barrier embryo implantation laboratory must enter through the sterile corridor and are required to shower and don sterile working attire. Following embryo implantation the female will be transferred to the small barrier animal housing room (brown - red arrow) for continuation of the pregnancy, parturition, and raising of the pups. At the time of weaning the female will be euthanized and a complete necropsy with extensive serology, microbiologic culture, and PCR testing performed to confirm that all pathogens have been excluded. Following satisfactory test results the pups would then be transferred to one of the two larger barrier animal housing rooms for continued breeding and colony expansion.