 The
Facility is designed to accommodate the culture of mammalian cell lines and
other animal cell lines, such as insect cells.
Work involving the culture of bacterial cells, spore-forming yeast, Neurospora
or fungi or single-celled microorganisms may not be performed due to the
potential of these cultures contaminating other cell lines.
All work involving human cell lines must first be approved by the University
Biosafety Committee. All work involving recombinant DNA must first be approved
by the University Recombinant DNA Committee. All work is reviewed for
appropriateness of the work for the Facility. The Principal Investigator submits
a request to perform work/proposal of the work to the Director. If special
approvals are needed as described above, these approvals must be submitted at
the time of the request to perform work.
Equipment and Supplies Provided by the Cell Culture Facility
The Cell Culture Facility is a BSL 2 Laboratory. All work will be performed
using BSL 2 guidelines. The facility is equipped with three Biological Safety
Cabinets.
Room 3314C has a BSL 2+ cabinet. Primarily work needing a higher level of safety
(such as viral work) will be performed in this room. A CO2 incubator is also
located in this room.
Room 3314A and 3314B both have a BSL 2 safety cabinet and a CO2 incubator.
Rainin LTS Pipettes-Dedicated for each hood
Pipet Controller-Dedicated for each hood
MVE Cryogenic storage system-for small scale storage. Contact director for large
scale freezing needs
Zeiss Inverted Microscope
37 deg water bath
Tabletop refrigerated centrifuge
Vortex
paper towels
disinfectants
kimwipes
biohazard bags
Other equipment may be purchased as the need arises.
Supplies provided by user
All other supplies needed including
Pipette tips—sterile and filtered
Plastic serological pipettes plugged
Plastic aspiration pipettes or plastic transfer pipettes
Culture flasks
Cryovials
Cell scrapers
Media
Gloves
Lab coats
Goggles
Masks
Antibiotics and other additives
15 ml and 50 ml conical tubes (disposable)
Any other miscellaneous supplies needed
BSL 2 Microbiological Practices
The door to the facility must be kept locked at all times. Check the door after
entering and leaving. Do not prop the door open.
Wash hands after handling viable material, after removing gloves and before
leaving the laboratory.
An eyewash station is available by the sink.
Eating, drinking, smoking handling contact lenses and applying cosmetics are
prohibited in the laboratory.
Mouth pipetting is prohibited. Use mechanical pipetting devices
All sharps must be placed in the sharps container
Work surfaces must be decontaminated upon completion of work and after any spill
or splash of viable material. Clean
up all spills. The hood is to be sterilized. Discide is provided for infectious
samples. It needs to be wiped off after one minute. Follow by spraying the walls
and the floor of the hood with 70% isopropanol. Noninfectious samples may use
only the 70% isopropanol. If a spill occurs in the incubator, the shelves must
be removed, washed and sterilized. If a culture dish has spilled, it must be
wrapped and placed in the biohazard waste. Culture flasks may be wiped with 70%
isopropanol and returned to the incubator. Report all spills outside the hood to
the director. Spills outside the hood are to be cleaned with 10% bleach or 70%
isopropanol depending on volume and area of spill (Example: Floor spills-clean
with bleach. Microscope spills-clean with isopropanol). All materials used in
decontaminating must be placed in the biohazard autoclavable waste bags.
If a spill is suspected inside contained equipment, such as a centrifuge, wait 5
minutes after the centrifuge has stopped to allow aerosols to settle. A HEP A
filtered face mask, gloves, lab coat and shoe protectors should be worn to clean
the spill. Use absorbent paper towels to absorb the spill, working from the
edges towards the center. Swab the surface with a disinfectant. At the end of
the clean-up, disposable protective clothing should be added to the waste bag
and autoclaved.
All solid waste must be autoclaved and placed in the biohazard trash boxes.
Liquid waste is to be decontaminated in 10% bleach for at leash 20 minutes
before discarding down the sink.
Report all insect and rodent problems to the director.
Animals are not permitted in the laboratory
Broken glassware must not be handled directly by hand. A dustpan and brush Is
located for your use. All broken glassware must be decontaminated before
disposal.
All cultures, specimens, or potentially infectious wastes must be placed in a
container with a cover that prevents leakage during collection, handling
processing, storage, transport, or shipping.
Safety Equipment (Primary Barriers)
Procedures with a potential for creating infectious aerosols or splashes are
conducted in the biological safety cabinet.
Face protection (goggles, mask, face shield or other splatter guard) must be
used for anticipated splashed or sprays of infectious or other hazardous
material to the face when the microorganisms must be manipulated outside the
biological safety cabinet.
A Protective laboratory coat and gloves are to be worn when performing any work
involving samples. The lab coat is to remain in the facility. Soiled lab coats
are to be disposed of in the biohazard waste. Gloves are to be disposed of when
overtly contaminated, and removed when work with infectious materials is
completed or when the integrity of the glove is compromised. Disposable gloves
are not washed, reused, or used for touching “clean” surfaces (keyboards,
telephones, etc.). They are not to be worn outside the lab. Hands are to be
washed following removal of gloves.
The biosafety cabinet provides additional protection. Loss of adequate airflow
in the BSL 2+ biological safety cabinet will trigger an alarm on the cabinet.
All work must immediately be terminated. The worker must immediately close all
open containers. The worker should remove all protective outer clothing and exit
the room, leaving the door to the BSL 2+ room closed. The loss of adequate
airflow must be reported to the director immediately upon exiting.
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