1.What is cell culture?
Cell culture refers to removing cells from animals or plants and then growing them in a favorable artificial environment. The cells can be taken directly from the tissue and broken down by enzymatic or mechanical means before culturing, or they can be derived from established cell lines or cell lines.
2.What is primary culture?
Primary culture refers to the culture stage after cells are separated from the tissue and proliferate under appropriate conditions until they occupy all available substrates (that is, reach confluence). At this stage, the cells must be subcultured by transferring them to a new container with fresh growth medium to provide more space for continued growth.
2.1 Cell line
After the first subculture, the primary culture is called a cell line or subclone. Cell lines derived from primary cultures have a limited lifespan (ie they are limited; see below), and as they pass, the cells with the highest growth capacity dominate, resulting in a certain degree of genotype in the population onsistent with phenotype.
2.2 Cell strain
If a subpopulation of a cell line is positively selected from the culture by cloning or some other method, the cell line would become a cell strain. Cell strains usually acquire additional genetic changes after the parental line starts.
3.Limited and continuous cell lines
Normal cells usually divide only a limited number of times before losing the ability to proliferate. This is a genetically determined event called senescence; these cell lines are called finite cell lines. However, some cell lines become immortal through a process called transformation, which can occur spontaneously or can be induced by chemicals or viruses. When a finite cell line undergoes transformation and gains the ability to divide indefinitely, it becomes a continuous cell line.
4.Culture condition
The culture conditions of each cell type are very different, but the artificial environment for culturing cells is always composed of a suitable container, which contains the following:
4.1 Substrate or culture medium that provides essential nutrients (amino acids, carbohydrates, vitamins, minerals)
4.2 Growth factors
4.3 Hormones
4.4 Gases (O2, CO2)
4.5 Regulated physical and chemical environment (pH, osmotic pressure, temperature)
Most cells are anchorage-dependent and must be cultured on a solid or semi-solid substrate (adherent or monolayer culture), while other cells can grow floating in the medium (suspension culture).
5.Cryopreservation
If there are excess cells in the subculture, they should be treated with an appropriate protective agent (such as DMSO or glycerol) and stored at a temperature below -130°C (cryopreservation) until they are needed. For more information about subculture and cryopreservation of cells.
6.Morphology of cells in culture
Cells in culture can be divided into three basic categories based on their shape and appearance (ie morphology).
6.1 Fibroblasts cells are bipolar or multipolar, have an elongated shape, and grow attached to the substrate.
6.2 Epithelial-like cells are polygonal, have a more regular size, and are attached to the matrix in discrete sheets.
6.3 Lymphoblast-like cells are spherical and usually grow in suspension without attaching to the surface.
7.Application of cell culture
Cell culture is one of the main tools used in cell and molecular biology. It provides an excellent model system for studying the normal physiology and biochemistry of cells (such as metabolic research, aging), the effects of drugs and toxic compounds on cells, and mutagenesis and carcinogenic effects. It is also used for drug screening and development and large-scale manufacturing of biological compounds (such as vaccines, therapeutic proteins). The main advantage of using cell culture for any of these applications is the consistency and reproducibility of the results that can be obtained using a batch of cloned cells.
Post time: Jun-03-2019