Cell technology animal cell large-scale culture method
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Animal cell culture can be carried out using three main methods: adherent culture, suspension culture, and immobilization culture, depending on the type of cells being grown. These techniques are essential for large-scale production in biotechnology and pharmaceutical industries.
One key aspect of animal cell culture is understanding their growth characteristics. Animal cells grow slowly, are prone to contamination, and often require antibiotics in the culture medium. They lack a cell wall, making them more fragile and sensitive to mechanical stress. Additionally, they have low tolerance for strong agitation and ventilation. Most animal cells exhibit anchorage dependence, meaning they must attach to a surface to grow. This also means that their growth is limited to about 50 generations before they degenerate. The products from these cultures are often found both inside and outside the cells, which increases the cost of production.
In vitro, animal cells can be classified into two types based on their attachment requirements: adherent-dependent and non-adherent-dependent. Adherent-dependent cells need to attach to a solid or semi-solid surface with some charge to grow, such as most non-lymphoid tissue cells. Non-adherent-dependent cells, like blood and lymphoid cells, do not require attachment and can grow freely in suspension.
The optimal temperature for most animal cell cultures is around 35–37°C, which matches the normal body temperature of mammals. Deviations from this range can significantly affect metabolism and survival. Cells generally tolerate lower temperatures better than higher ones. At 39–40°C, cells may suffer damage but can recover, while above 43°C, many cells die. At lower temperatures (30–20°C), metabolism slows down, leading to reduced nutrient exchange and eventual detachment from the substrate.
Adherent culture involves growing cells on a solid surface. This method allows for easy media changes and perfusion, and some cells produce higher yields when attached. However, it requires more space and is harder to scale up compared to suspension culture. Common systems include rotating bottles, hollow fibers, and microcarriers.
Suspension culture, on the other hand, is ideal for non-adherent cells such as hybridoma cells. It mimics microbial fermentation and is easier to scale up. Serum-free suspension culture is gaining popularity because it reduces purification steps and improves product quality.
Immobilization culture involves attaching cells to a carrier material, which helps protect them from shear stress and makes product recovery easier. Methods include adsorption, covalent bonding, cross-linking, embedding, and microencapsulation. Each has its own advantages and limitations, such as ease of use, cell retention, and diffusion issues.
Apoptosis is a major cause of cell death in large-scale cultures, with up to 80% of cell loss attributed to programmed cell death rather than necrosis. Caspases play a central role in this process. Strategies to combat apoptosis include adding nutrients like glutamine, manipulating genes such as Bcl-2, and using chemical inhibitors to block apoptotic pathways.
By addressing these challenges, researchers can enhance cell viability, increase productivity, and improve the efficiency of large-scale animal cell culture processes.