Essential Laboratory
Equipment for Cell Culture
Dr. Khalid Enan
DFG-Sponsored Workshop, Assuit University, Egypt
10 Sep 2012
Definition of cell culture :
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Cell culture refers to the culturing of cells derived
from multicellular eukaryotes, especially animal
cells
A complex process by which cells are grown under
controlled conditions
Used to
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diagnose infections
test new drugs
in research.
Basic requirements for cell
culture
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Sterile work area
Laminar flow hood
Biohazard cabinet
Sterile work area:
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A separate room should be made available
for clean cell culture work.
This room should be free of through traffic
and equipped with an air flow cabinet,
which supplies filtered air around the work
surface.
Cell culture room
Laminar flow hood
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Strict sterility is needed.
Laminar flow hood offers the best sterile
protection.
Suitable only for preparing medium and
other sterile reagents and for culturing nonprimate cells.
Laminar-Flow Hood
Biohazard cabinet:
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Used for potentially hazardous materials
including cell lines, cultures for virus
propagation, or toxic drugs).
Biohazard cabinet
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All work surfaces, benches and shelves and
the base of the airflow cabinets must be
kept clean by frequent swabbing with 70%
ethanol or an alternative disinfectant.
Incubation facilities:
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The cell culture laboratory will need to be
furnished with an incubator or hot room to
maintain the cells at 30-40 °C. The incubation
temperature will depend on the type of cells being
cultivated.
It may be necessary to use an incubator which has
been designed to allow CO2 to be supplied from
a main supply or gas cylinder so that an
atmosphere between 2-5% CO2 is maintained in
the incubator
Incubators
Fridges and freezer (-20 °C)
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Both items are very important for storage of
liquid media at 4 °C and for enzymes (e.g.
trypsin) and some media components (e.g.
glutamine and serum) at -20 °C.
A fridge or cold room is required to store medium
and buffers. A freezer will be needed for keeping
pre-aliquoted stocks of serum, nutrients and
antibiotics. Reagents may be stored at -20 °C.
For prolonged preservation, cells may be
preserved in liquid nitrogen or a -70 °C freezer.
Fridges and freezer (-20 °C)
Microscopes
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A simple inverted microscope is essential
so that cultures can be examined in flasks
and dishes.
A very simple light microscope with x100
magnification for routine cell counts in a
hemocytometer.
Inverted Microscope
Tissue culture ware (Flasks)
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A variety of tissue culture plastic ware is
available, the most common being
specially treated polystyrene.
Cells can be maintained in flasks (25 cm2
or 75 cm2), tissue culture tube, microtitre
plates………
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Filter sterilization
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Non-autoclavable substances, such as
culture media, enzymes, hormones, and
bicarbonate buffers, must be sterilized
through a 0.22 μm pore size membrane
filter. These are obtainable in various
designs to allow a wide range of volumes
to be filtered (e.g., Millipore, Gelman).
Filter sterilization:
Liquid N2 System
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It is important to maintain continuity in
cells to prevent genetic drift and to guard
against loss of the cell line through
contamination and other disasters.
Facilities for counting cells
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It is possible to monitor cell growth by
eyes (looking for confluence), however,
more accurate cell counts are required for
most experimental purposes.
The most commonly used device is the
Improved Neubauer hemocytometer
originally designed for counting blood
cells.
Facilities for counting cells
Further required equipment:
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Centrifuge
Centrifuge tubes and universal containers
Water bath
Vacuum pump
Graduated pipettes of various sizes.
Disposable Pasteur pipettes.
Rubber bulbs or pipette-aid
Automatic pipettes (volumes of 1-1000 μl).
Plastic tips.
Graduated glass or plastic cylinders.
water bath
vacuum pump
graduated pipettes
centrifuge tube (Falcon tube)
Pasteur pipettes.
Eppendorf tube
rubber bulbs
automatic pipettes
Pipet-aid
Centrifuge
Question ?
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