Animal cell culture involves growing animal cells outside their natural environment under controlled laboratory conditions.
This process of Growth of Animal Cells in Culture allows for the study of cell biology, physiology, and biochemistry in a controlled setting, facilitating research and development in various biomedical fields.
General Procedure for Cell Culture
Preparation of Culture Media:
Nutrient-rich media are prepared, containing essential amino acids, vitamins, minerals, glucose, and growth factors.
Serum (like fetal bovine serum) may be added to provide additional growth factors.
Sterilization and Aseptic Techniques:
All equipment and media are sterilized to prevent contamination.
Work is conducted in a laminar flow hood to maintain a sterile environment.
Isolation of Cells:
From Tissue: Cells are isolated from animal tissues using enzymatic digestion (e.g., with trypsin or collagenase) to separate cells.
From Established Lines: Cells are obtained from existing cell lines stored in repositories.
Seeding Cells:
The isolated cells are placed into culture vessels (flasks, dishes) containing the prepared media.
Incubation:
Cultures are incubated at optimal conditions (typically 37°C with 5% CO₂) to support cell growth and function.
Monitoring and Maintenance:
Regular observation under a microscope to assess cell health and confluency.
Periodic media changes to supply fresh nutrients and remove waste products.
Subculturing (Passaging):
When cells reach sufficient growth (confluency), they are transferred to new vessels to continue proliferation.
Types of Cell Cultures (Growth of Animal Cells)
Primary Cell Cultures:
Derived directly from animal tissues.
Have a limited lifespan (finite number of divisions).
Closely resemble the physiological state of cells in the body.
Applications: Studying normal cell functions, drug metabolism, and toxicology.
Established (Continuous) Cell Lines:
Cells that have adapted to in vitro conditions and can proliferate indefinitely.
May have undergone genetic changes to become immortalized.
Examples: HeLa cells, HEK 293 cells.
Applications: Large-scale studies, vaccine production, recombinant protein expression.
Transformed Cell Cultures:
Cells that have been genetically altered to exhibit uncontrolled growth (similar to cancer cells).
Often used to study cancer mechanisms and test anti-cancer drugs.
Characteristics: Loss of contact inhibition, anchorage-independent growth.