ABP Human Cells in Rodents

1. Purpose and Scope

This Animal Biosafety Procedure (ABP) describes prudent practices, procedures, and equipment to reduce risk when implanting human cells in rodents at Animal Biosafety Level 2 (ABSL-2). Potential hazards include bloodborne pathogens (BBPs) such as HIV, Hepatitis B, Hepatitis C, and other human bloodborne pathogens. It is important to recognize that BBPs are not limited to HIV and Hepatitis viruses; other pathogens, such as syphilis, malaria, and certain arboviruses, can also pose risks. The practices and procedures outlined in this document are in accordance with those described in the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL), 6th edition, and conform to the Occupational Safety and Health Administration (OSHA) Bloodborne Pathogen Standard, 29 CFR 1910.1030.

2. Responsibilities

    Principal Investigator (PI): Ensures that personnel are aware of the hazards associated with human cells and that they receive appropriate training before starting ABSL-2 experiments. Personnel must comply with the safe work practices and procedures described in this ABP.
   Personnel: Must follow the safe work practices and procedures outlined in this ABP

3. Administrative Controls

• 3.1 Training
  • 3.1.1 Receive laboratory-specific training for manipulation of human cells.
  • 3.1.2 Complete annual OSHA training for bloodborne pathogens and review the laboratory’s Exposure Control Plan.
  • 3.1.3 Complete the ABSL-2 online training module and CARE training for handling of rodents.
  • 3.1.4 CARE and EHS will provide additional on-site training, as necessary.
• 3.2 Access and Signage
  • 3.2.1 Inform the facility manager prior to using human cells in rodents.
  • 3.2.2 Review the hazards and potential risks of the experiment, and complete IACUC module 2 before accessing the animal facility.
  • 3.2.3 The facility supervisor will post a hazard sign at the animal room. Research, EHS, and CARE staff will develop information contained in the sign, which will include:
    • The biohazard symbol and ABSL-2 designation
    • Human cells
    • Personal protective equipment
    • Disinfectant(s)
    • Contact information for CARE, EHS, and Cornell Health
  • 3.2.4 Post a hazard ID card on cages that contain animals implanted with human cells. The card will include the biohazard symbol, type of cells, and date of implantation.
• 3.3 Medical Surveillance
  • 3.3.1 Participate in the Animal Users Health and Safety Program (AUHSP).

4. Work Practice and Procedure Controls

• 4.1 Implantation of Cells
  • 4.1.1 See Sections 5.1, engineering controls, and 6.1, personal protective equipment (PPE).
  • 4.1.2 If cells have been previously passaged through rodents, they should be screened for potential rodent pathogens before continued use. Refer to Animal Care and Use Procedure 619, Tumor and Cell Line Testing
  • 4.1.3 Use an appropriate manual or physical restraint device. If the procedure or conditions of implantation pose too high a risk with an awake animal (e.g., cranial injections, inexperienced individual performing the procedure), sedate the animal prior to implantation.
  • 4.1.4 Use a disinfectant-soaked cloth to wipe away excess inoculum leaking from inoculation site.
• 4.2 Sharps Handling
  • 4.2.1 Substitute plasticware for glassware whenever possible, and implement the following practices described in the table:

    • Safe Practices for Handling Sharps
      Limit the use of sharps to when no other alternatives are available
      Keep all sharps in full view at all times
      Use only Luer-lock syringes and needles or units where the needle is integral to the syringe
      Implement safety engineered sharps where practical (retractable needles, needle tip shields, self-sheathing scalpels, etc)
      Dispose of sharps directly, without manipulation (i.e., do not bend, shear, break, recap, or use hands to remove needles from syringes or blades from scalpels), in an approved sharps disposal container. Maintain disposal container in animal room within arm’s reach
      Handle broken glass or other sharps with a secondary device such as forceps or broom and dustpan- not your hands

  • 4.2.2 Do not recap needles. 
    • If recapping must be done: first receive approval by EHS and the IACUC. Once approved, only use one of the following two methods: one handed scoop technique; forceps or tongs to place the cap on the needle.
  • 4.2.3 If a needle/scalpel must be set down temporarily during a procedure, set the sharp end inside a clean 50-mL conical tube or equivalent to act as a protective sheath (rather than recapping) and to prevent injury.
• 4.3 Hygiene
  • 4.3.1 Eating, drinking, smoking, handling contact lenses, applying cosmetics, storing food for human consumption, and mouth pipetting are strictly prohibited in animal facilities.
  • 4.3.2 Wash hands thoroughly with soap and water after removing gloves. Use an alcohol-based hand sanitizer if sink is not readily available.
  • 4.3.3 If working long hours in a rodent room consider taking a full body shower to reduce the amount of potential allergens present on your body.
    • For more information, see ACUP 713: Hygiene – Hand Washing
• 4.4 Decontamination and Spill Response
  • 4.4.1 Decontaminate work surfaces and equipment (e.g., inside of biosafety cabinet, animal cages) with an EPA-registered disinfectant for human bloodborne pathogens - allow at least 5-10 minutes of contact time.
  • 4.4.2 Cover spills with absorbent towels/pads and saturate with disinfectant. Allow 5-10 minutes contact time to achieve adequate disinfection. Appropriately segregate waste in red biohazard bags or sharps disposal containers and re-apply disinfectant to spill area.
• 4.5 Handling of Waste
  • 4.5.1 Inside a biosafety cabinet, place a water soaked paper towel in the dirty cage, to generate steam when autoclaved, or leave water bottle in cage. Wipe exterior of cage with disinfectant before removing from biosafety cabinet.
  • 4.5.2 Dispose of sharps-related items (e.g., needles, syringes, Pasteur pipettes, blood tubes) directly in a sharps disposal container.
  • 4.5.3 Dispose of non-sharps items (e.g., gloves, intact plasticware) in a red biohazard bag.
  • 4.5.4 Treat infectious liquid waste with concentrated household bleach to a final volume of 10% bleach and allow at least 30 minutes contact time before disposal in the sanitary waste drain- follow with copious amounts of water.
  • 4.5.5 Place carcasses (no gloves, plastic, etc.) in compostable bags suitable for disposal in the Waste Management digester. Wipe bags with appropriate disinfectant and store all bags in a larger biohazard bag or biohazard-labeled drawer in refrigerator. Alternatively, roll up carcasses in bench diapers and place them directly in biohazard bags.
  • 4.5.6 Animal care staff will dispose of waste and carcasses, unless other arrangements are made.
• 4.6 Transport of Biohazardous Materials
  • 4.6.1 Transport infectious agents and contaminated samples between the laboratory and animal facility in a sealed, secondary container with absorbent toweling, and labeled with the biohazard symbol. The container should be drop proof.
• 4.7 Tissue Harvest
  • 4.7.1 Perform tissue harvest in a certified class II biosafety cabinet. Collect fluids using a tray or bench diaper. Preferably, secure the carcass with tape instead of pins.
  • 4.7.2 When possible use only one sharps item (e.g., scalpel, scissors) at a time and keep in full view.
  • 4.7.3 Place any harvested tissue or fluids in appropriate primary containers (e.g., screw top vial, sealable plastic bag), decontaminate exterior, and transport as per section 4.6. Fixed tissues (e.g., 10% buffered formalin) are no longer considered biohazardous. Use appropriate personal protective equipment when handling these samples and transport in a secondary container.
  • 4.7.4 Follow the sharps handling practices outlined in section 4.2.

5. Engineering Controls

• 5.1 Use of Biosafety Cabinet
  • 5.1.1 Perform all procedures carefully to minimize the creation of aerosols. Use a certified class II biosafety cabinet for: implantation; necropsy and tissue harvest; and cage changing.
  • 5.1.2 Wipe cages with appropriate disinfectant when moving out of biosafety cabinet.
• 5.2 Housing and Handling of Animals
  • 5.2.1 House animals in a primary containment device appropriate for the rodent species, such as a ventilated micro-isolator cage or static micro-isolator cage with a filter top.
  • 5.2.2 Conduct implantations, cage changing, and other procedures in a biosafety cabinet.
  • 5.2.3 Whenever possible, use forceps to transfer animals between cages.

6. Personal Protective Equipment (PPE)

• 6.1 Don the following minimum PPE before entering ABSL-2 animal rooms:
  • Disposable fluid resistant  gown
  • Disposable gloves (nitrile- avoid latex when possible) - Use double gloves when handling human cells or when implanting animals. Outer glove should overlay cuff of gown
  • Shoe covers
• 6.2 Wear additional PPE (e.g., face shield, respiratory protection, cut/bite resistant gloves) when appropriate, engineering controls are not available, or as indicated by the vector, hazards, or experimental conditions.
• 6.3 Solid toed shoes are required for entry into animal rooms.
• 6.4 Change gloves frequently (or decontaminate with disinfectant) during activities to avoid contamination of equipment and surfaces. Remove and replace other PPE if contaminated or breached.
• 6.5 Remove PPE before exiting the animal room and dispose in red biohazard bag. First remove outer gloves, gown (turning inside out), shoe covers while stepping out of the room (step-over technique), and finally inner gloves.
  • For more information, see ACUP 715: Personal Protective Equipment

7. Response to Accidental Exposures

• 7.1 Personnel who sustain an overt exposure such as a splash to mucous membranes, direct contact with open wounds, or a sharps injury should:
  • Wash exposed area with soap and water or rinse in eye wash for at least 5 minutes
  • Perform first aid, if applicable
  • Notify supervisor
  • Seek medical evaluation at a medical provider as soon as possible after an exposure. Have SDS or other information document readily available. After hours seek evaluation at Cayuga Medical Center.
  • Document exposures, injuries, and illnesses in the Cornell University Injury/Illness/Exposure Report, Incident Reporting
  • For more information, see ACUP 707: Animal Related Injury

8. Emergency Phone Numbers

• Police, Fire, and Medical Emergencies: calling 911 from an on-campus phone or by calling 607-255-1111 from a cellular phone
• Environmental Health & Safety (EHS): 607-255-8200 (off hours 607-255-1111)
• Cornell Animal Resources and Education (CARE): 607-253-4378 (off hours 1-800-349-2456 for veterinary medical emergencies)

Reviewed July 1, 2024