Cell Culture Lab Practices Quiz

Cell Culture Lab Equipment & Supplies

• Basic cell culture lab equipment and supplies are crucial for successful cell culture experiments.

Incubators

• Incubators provide the optimal environment for cell growth.
• Stainless steel incubators are easy to clean and resist corrosion, especially if humid air is needed.
• Frequent cleaning prevents contamination.

Types of Incubators

• Dry incubators: More economical but require sealed flasks to prevent evaporation. A water dish can help with humidity, but precise control is not possible.
• Humid CO₂ incubators: More expensive but offer superior control over culture conditions. They are used with Petri dishes or multi-well plates needing a controlled atmosphere with high humidity and increased CO₂ tension.

Storage

• Cell culture labs need storage areas for liquids like media and reagents, chemicals like drugs and antibiotics, consumables (pipettes, gloves), glassware (media bottles, pipettes), specialized equipment, and tissues/cells.

Types of Storage

• Refrigerators: For small labs, a domestic refrigerator is adequate to store reagents/media at 2-8°C. Larger labs often use cold rooms for cell cultures. A domestic freezer can be a cheaper alternative to a dedicated lab freezer.
• Freezers: Most cell culture reagents can be stored at -5°C to -20°C. An ultradeep freezer (-80°C) is sometimes used, but not always necessary. Some reagents (antibiotics, enzymes) require freezers without autodefrost cycles.

Cryogenic Storage

• Cell lines in continuous culture can suffer from genetic instability as their passage number increases. Cryogenic storage is essential for preparing working stocks of the cells.
• Do not store cells in -20°C or -80°C freezers; their viability quickly decreases when stored at these temperatures.

Liquid Nitrogen

• Liquid nitrogen storage systems come in vapor phase and liquid phase.
• Vapor phase systems decrease explosion risk in cryostorage tubes and are needed for biohazardous materials.
• Liquid phase systems provide longer holding times and are often more economical.

Cell Counter

• Cell counters are used to quantify cell growth and kinetics.
• They count cells and calculate cell viability.
• Data presentation is visual, and data is securely stored.
• Data can be exported for further analysis.

Hemocytometer

• A glass slide with a central chamber grid to count cells under a microscope.
• Viability is determined using trypan blue exclusion, where viable cells don't take up the dye.

Cell Culture Flask & Plate

• Used for cell cultivation.

Multichannel Pipette

• Automated pipette to add liquids to multiple samples simultaneously.

Buffer

• Important solutions used in cell culture (example, PBS).

Centrifuge

• Used to separate or concentrate cells/cell components.
• Precise temperature control and centrifugal speeds are vital.

Laminar Flow

• Protects the working environment from contaminants. HEPA-filtered air is constantly supplied in a unidirectional flow. Different designs provide protection to the culture or the user.

Cell Culture Lab Safety

• Follow specific safety protocols to prevent contamination.

Safety Protocols

• Handwashing: Always wash hands thoroughly.
• Decontaminating work surfaces: Clean and disinfect surfaces to prevent contamination.
• Handling carcinogens: Handle carcinogenic materials with care using protective equipment.
• Quarantine new cells: Isolate new cells to prevent contamination of existing cell cultures.
• Screen existing cell stocks: Regularly test for contamination in existing cell cultures.

Biohazards in Cell Culture

• Viruses and modified cells are significant possible biohazards.
• Use and generation of modified cells can pose risks.
• Workers must never culture cells from their own body tissues.

GMOs (Genetically Modified Organisms)

• Genetically modified organisms (GMOs) should be strictly controlled and regulated.
• Most countries have regulatory organizations to minimize risks posed by GMOs.
• UK examples: require a Genetic Modification Safety Committee (GMSC) for labs working with GMOs.

Cell Culture Waste Disposal

• Inactivate tissue culture waste (like culture media) with hypochlorite for at least 2 hours before disposal to drain with excess water.
• Contaminated pipettes should be put into overnight hypochlorite solution before incineration.
• Solid waste (flasks, tubes, gloves, etc.) should be placed in heavy-duty sacks for incineration. Autoclaving is sometimes a less desirable treatment method.
• Waste from genetically modified (GMO level 2) organisms often requires specialized treatment and tracking.