Aseptic Techniques: The facility must maintain sterile conditions to prevent contamination by microorganisms (bacteria, fungi, viruses). This includes using sterile equipment, reagents, and culture media.
Temperature: Cells are usually grown at specific temperatures (typically around 37°C for mammalian cells) that mimic their natural environment. Some cells require lower or higher temperatures.
Humidity: Maintaining appropriate humidity levels (around 95%) is essential to prevent evaporation of culture medium.
CO2 Concentration: A regulated CO2 environment (usually 5%) is critical to maintain the pH of the culture medium, especially when using bicarbonate-based buffers.
Oxygen Levels: For some cell types, oxygen concentration is controlled to simulate physiological conditions.
Light: Cells are typically grown in the dark or under controlled light to avoid photo-damage.
Subculturing: Cells need to be subcultured or passaged periodically to prevent overcrowding and to ensure they continue to grow healthily.
Cryopreservation: Cell lines may be frozen for long-term storage using cryoprotectants like dimethyl sulfoxide (DMSO) to preserve cell viability.
Monitoring Growth: The growth rate of cells is monitored regularly to determine their health, confluence, and viability.
Culture Media: Cells are grown in a nutrient-rich medium that provides essential growth factors, amino acids, vitamins, and minerals. The composition of this medium must be tailored to the specific needs of the cells.
Feeding: Medium is changed periodically to provide fresh nutrients and remove metabolic waste products.
pH and Osmolarity: Maintaining the correct pH (usually around 7.4) and osmolarity is essential for the health of the cells.
Primary Cells: Directly isolated from tissues and more closely resemble the natural state of cells. They may require specific conditions or growth factors.
Immortalized Cell Lines: Cells that have been genetically modified to divide indefinitely, making them easier to maintain and culture for long periods.
Stem Cells: Require special culture conditions to maintain pluripotency or induce differentiation.
Contamination Monitoring: Regular tests for bacterial, fungal, and viral contamination are vital to maintaining the purity of cell cultures.
Cell Authentication: Regular cell line validation is required to confirm that the correct cell type is being used and that it hasn’t been misidentified or cross-contaminated.
Environmental Monitoring: The environment must be regularly assessed for temperature, humidity, CO2 levels, and air quality to ensure compliance with optimal culture conditions.
Biosafety: Adequate biosafety measures must be in place to handle hazardous biological materials. This includes the use of biosafety cabinets, proper disposal of biological waste, and protective gear for personnel.
Regulations: Facilities must comply with local and international regulations, such as Good Manufacturing Practices (GMP), Good Laboratory Practices (GLP), and institutional review boards (IRB), especially when working with human cells or tissues.
Robotic Systems: Automated systems can assist with tasks like cell counting, media exchange, and cryopreservation, ensuring consistency and reducing the risk of human error.
Monitoring Systems: Use of sensors and data collection devices to monitor culture conditions in real time, such as pH, temperature, dissolved oxygen, and cell density.
When using human cells, stem cells, or animal-derived cells, ethical guidelines and proper consent processes must be followed. This includes sourcing cells responsibly and ensuring they are obtained in compliance with ethical standards.
the core principles of a cell culture facility revolve around maintaining sterile, controlled, and optimized environments for cell growth while adhering to safety, regulatory, and ethical standards.