Implantation of Human Cells in Laboratory Rodents

  1. Purpose and Scope

This Animal Biosafety Procedure (ABP) describes prudent practices, procedures, and equipment to reduce riskwhenimplanting human cells in rodents at Animal Biosafety Level 2 (ABSL-2). Potential hazardsinclude bloodborne pathogens such as HIV, Hepatitis B and C. The practices and procedures outlined in this documentare in accordance with those described in the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th edition ( and conform to the Occupational Safety and Health Administration (OSHA) Bloodborne Pathogen Standard, 29 CFR 1910.1030.

***Alternative practices, procedures, and equipment may be used, but they must be described in a user-generated standard operating procedure, and approved by EHS and the IACUC before use.***

  1. Responsibilities

The Principal Investigator will ensure that personnel are made aware of the hazards associated with human cells and that they receive training commensurate with their activities prior to commencing ABSL-2 experiments. Personnel will comply with the safe work practices and procedures described within this Animal Biosafety Procedure.

  1. Administrative Controls
  2. Training
  3. Receive laboratory-specific training for manipulation of human cells.
  4. Complete annual OSHA trainingfor bloodborne pathogens and review the laboratory’s Exposure Control Plan.
  5. Complete the ABSL-2 online training module and CARE trainingfor handling of rodents.
  6. CARE and EHS will provide additional on-site training, as necessary.

3.2Access and Signage

3.2.1Inform the facility manager prior to using human cells in rodents.

3.2.2Review the hazards and potential risks of the experiment, and complete IACUC module 2 before accessing the animal facility.

3.2.3The 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
  • The name of the Principal Investigator and IACUC protocol number
  • Human cells
  • Personal protective equipment
  • Disinfectant(s)
  • Contact information for CARE, EHS, and Gannett Health Services
  • Post a hazard ID card on cages that contain animals implanted with human cells. The card will includethe biohazard symbol, type of cells, and date of implantation.

3.3Medical Surveillance

3.3.1Participate in the Animal Users Health and Safety Program (AUHSP).

  1. Work Practice and Procedure Controls
  2. Implantation of Cells
  3. See Sections 5.1, engineering controls, and 6.1, personal protective equipment (PPE).
  4. If cellshave 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 (
  5. 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 toimplantation.
  6. Use a disinfectant-soaked cloth to wipe away excess inoculum leaking from inoculation site.

4.2Sharps Handling

4.2.1Substitute plasticware for glassware whenever possible, andimplement the following 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
  • Dispose of sharps directly, without manipulation,in an approved sharps disposal container (i.e., do not bend, shear, break, recap, or use hands to remove needles from syringes or blades from scalpels). Maintain disposalcontainer within arm’s reach (including inside a biosafety cabinet)
  • Handle broken glass or other sharps with a secondary device such as forceps or broom and dustpan- not your hands
  • Do not recap needles. However, if recapping must be donefirstreceive approval by EHSandthe IACUC, and use one of the following two methods: one handed scoop technique; forceps or tongs to place the cap on the needle.

4.3Hygiene

4.3.1Eating, drinking, smoking, handling contact lenses, applying cosmetics, storing food for human consumption, and mouth pipetting are strictly prohibited in animal facilities.

4.3.2Wash hands thoroughly with soap and water after removing gloves. Use an alcohol-based hand sanitizer if sink is not readily available.

4.3.3If working long hours in a rodent room consider taking a full body shower to reduce the amount of potential allergens present on your body.

4.4Decontamination and Spill Response

4.4.1Decontaminate work surfaces and equipment (e.g., inside of biosafety cabinet, animal cages) with anEPA-registered disinfectant for human bloodborne pathogens ( - allow at least 5-10 minutes of contact time.

4.4.2Cover 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.

4.5Handling of Waste

4.5.1Inside 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.

4.5.2Dispose of sharps-related items (e.g., needles, syringes, Pasteur pipettes, blood tubes) directly in a sharps disposal container.

4.5.3Dispose of non-sharps items (e.g., gloves, intact plasticware) in a red biohazard bag.

4.5.4Treat 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.

4.5.5Place carcasses (no gloves, plastic, etc.) in 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. These bags will be disposed of offsite.

4.5.6Animal care staff will dispose of waste and carcasses, unless other arrangements are made.

4.6Transport of Biohazardous Materials

4.6.1Transport 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.

4.7Tissue Harvest

4.7.1Perform tissue harvest in a certified class II biosafety cabinet- use a tray or bench diaper to collect fluids. Use tape instead of pins to secure carcass.

4.7.2When possible use only one sharps item (e.g., scalpel, scissors) at a time and keep in full view.

4.7.3Place 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.

4.7.4Follow the sharps handling practices outlined in section 4.2.

  1. Engineering Controls
  2. Use of Biosafety Cabinet
  3. 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.
  4. Wipe cages with appropriate disinfectant when moving out of biosafety cabinet.

5.2Housing and Handling of Animals

5.2.1House 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.

5.2.2Conduct implantations, cage changing, and other procedures in a biosafety cabinet.

5.2.3Whenever possible, use forceps to transfer animals between cages.

  1. Personal Protective Equipment (PPE)

6.1Don the following minimum PPE before entering ABSL-2 animal rooms:

  • Disposable fluid resistant, solid front 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
  • Wear additional PPE (e.g., face shield, respiratory protection, cut/bite resistant gloves) when appropriate engineering controls are not available, or as indicatedby the vector, hazards, or experimental conditions.
  • Solid toed shoes are required for entry into animal rooms.
  • 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.
  • 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.
  1. Response to Accidental Exposures
  2. 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 10 minutes
  • Perform first aid, if applicable
  • Notify supervisor
  • Seek medical evaluation at Gannett Health Services, Occupational Medicine (255-6960) as soon as possible after an exposure. Have MSDS 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,
  1. Emergency Phone Numbers
  • Police, Fire, and Medical Emergencies: 911
  • Environmental Health & Safety (EHS): 255-8200 (off hours 255-1111)
  • Gannett Health Services, Occupational Medicine: 255-6960 (off hours 255-5155)
  • Cornell Animal Resources and Education (CARE): 253 4378 (off hours 1-800-349-2456 for veterinary medical emergencies)
  1. References

9.1Biosafety in Microbiological and Biomedical Laboratories, 5th edition. 2009. Centers for Disease Control and Prevention, National Institutes of Health.

9.2Bloodborne Pathogens Standard, 29 CFR 1910.1030. Occupational Safety and Health Administration.

9.3Working at Animal BSL 2. American Biological Safety Association.

Approved by: Institutional Biosafety Committee 12/13/11
Last revised by: Frank A. Cantone
Revision date: 10/31/11 / Animal_Biosafety_Procedures_Human_Cells_Rodents_v1.8
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