Essentially, cell culture involves the distribution of cells in an artificial environment (in vitro) which is composed of the necessary nutrients, ideal temperature, gases, pH and humidity to allow the cells to grow and proliferate.
Whereas pieces of tissue can be put in the appropriate culture to produce cells that can then be used for culture (explant culture), cells from tissues (soft tissue) can be obtained through enzymatic reactions. Such enzymes as trypsin and proname are used to break down the tissue and release the desired cells.
When cells have been obtained directly from the organism/animal tissue (or even plant tissue) through enzymatic or mechanical techniques, such cells are referred to as primary cells. However, cells that continue to proliferate indefinitely (after the first subculture) under special conditions are referred to as cell lines.
These particular cells tend to have been passaged for a long period of time, which causes them to acquire homogenous (similar) genotypic and phenotypical traits.
In addition, for safety purposes, work on cell culture should be carried out in the appropriate laminar flow hood, where air is directed away from the researcher.
Always check the information on the container to ensure that the medium is appropriate for the cell to be cultured,
Once prepared, the cell culture should be maintained under the recommended temperature range,
Monitor the culture every 30- 48 hours and check for confluency (when cells completely cover the surface of the culture) - However, this is largely dependent on the type of cells.
Once the procedure is completed and the cells have been analyzed, the culture should be appropriately discarded. Here, it's important to take a lot of caution given that by this time, cells have already proliferated and increased in numbers. Moreover, there are high chances that the specimen has been contaminated, which increase the risks of causing infections to the researcher if not handled appropriately.
In general, cell culture, whether it involves using a suspension or a stationary media, involves the growth of cells in an artificial environment with favorable conditions. Whereas enzymatic action can be used to obtain cells for culture, it's the mechanical disaggregation method that is most preferred given that it provides a simpler and less traumatic way of obtaining cells.
This method simply involves the slicing of a tissue into smaller pieces from which the spill out cells is then collected. Also, primary explants technique can be used to obtain the cells. However, this method is mostly useful for the disaggregation of smaller quantities of tissue.
While any cells can be used in cultures to observe their behavior, embryonic tissues are preferred (for primary cells) as compared to adult cells because they provide cells that are more viable and can rapidly proliferate.
Here, it is also important to ensure that that the cells are of higher quantity given that their survival rate tends to be lower in comparison to the sub-cultures. I
n order to enhance the success of cultures, it's also important that ensure that damage to the cells is minimized both during cell collection and processing. This is in addition to using the appropriate medium for the cells in question.
Along the same lines, take a look at types and techniques of tissue culture.
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As well as information on cell division, cell differentiation and cell staining and gain some insight into cell theory.
For the more advanced and a great read is Molecular Biology of the Cell. As well as learning about Cytochemistry.
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