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Will 3D cell culture methods have a future?
Primary cell culture is the disassociation of cells from a parental animal or plant tissue through enzymatic or mechanical measures and maintaining the growth of cells in a suitable substrate in glass or plastic containers under controlled environmental conditions. Cells in primary culture have the same karyotype (number and appearance of chromosomes in the nucleus of a eukaryotic cell) as those cells in the original tissue. Primary cell culture could be classified into two based on the kind of cells used in culture. Â· Anchorage Dependent or Adherent Cell â These cells require an attachment for growth. Adherent cells are usually derived from tissues of organs, for instance from kidney where the cells are immobile and embedded in connective tissue. Â· Anchorage Independent or Suspension Cells â These cells do not require an attachment for growth. In other words, these cells do not attach to the surface of culture vessel. All suspension cultures are derived from cells of the blood system; for instance, white blood cell lymphocyte is suspended in plasma. Cells derived from primary cultures have a limited life span. Cells cannot be held indefinitely due to several reasons. Increasing cell numbers in primary culture will lead to exhaustion of substrate and nutrients. Also, cellular activity will gradually increase the level of toxic metabolites in the culture inhibiting further cell growth. At this stage, a secondary or a subculture has to be performed to ensure continuous cell growth.
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Toxic metabolites such as alcohol, ammonia, ketones, amino acids, pesticides and polychlorinated biphenyls (PCB), can be released during growth, leading to the generation of free radicals. A secondary culture is needed to keep the cells healthy and prevent their elimination due to the toxic metabolites. The time taken for primary cell cultivation can range from ten minutes to several days. Following is the basic process of primary cell culture followed by a discussion on the possible adverse effects of certain drugs taken in primary culture. In general, all drugs taken in primary culture are metabolized by specific enzymes in the liver and kidney. Although the enzymes are present in all these tissues for all substances, most drugs are metabolized in their tissue of origin (e.g., liver; kidneys). The drug molecules that are toxic are generally removed from the system through various metabolic pathways following the action of the enzymes, which are.