Cell culture is the process by which cells are grown under controlled conditions, generally outside their natural environment. After the cells of interest have been. What is Cell Culture? Cell culture refers to the removal of cells from an animal or plant and their subsequent growth in a favoriable artificial environment. Cell culture is the technique in which cells are removed from an organism and placed in a fluid medium. Under proper conditions, the cells can live and even.
Open in a separate window. Growth conditions, cell morphology, and population in 2D and 3D cultures In traditional 2D monolayer culture, cells adhere and grow on a flat surface. The structures of 3D spheroids While various cell lines form nondistinct monolayers in 2D cell culture, distinctive differences in the structure of spheroids emerge as each cell line is cultured in 3D.
Cell receptor, protein, and gene expression in 3D cultures versus 2D culture In addition to the previously mentioned differences in physical and physiological properties, researchers have also found that cells in a 3D culture environment differ in gene, protein, and cell receptor expression from 2D-cultured cells.
Cellular characteristics 2D 3D Refs. Applications of 3D cell culture-based biosensors Although the versatility of 3D cell-based biosensors gives them a plethora of biomedical and bioanalytical applications, 91 including early detection and chronic management of illness 92 and environmental monitoring, 93 biosensors are prolific in pathogen testing, toxicology assays, and drug screening.
Conclusions It is increasingly evident that 3D cell culture models are better models than the traditional 2D monolayer culture due to improved cell—cell interactions, cell—ECM interactions, and cell populations and structures that resemble in vivo architecture.
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As such, they can be used for a number of purposes including;. On the other hand, culture may be categorized as;. Selective media - This is a special type of media that only allows for certain cells to grow.
As mentioned above, different types of synthetic media are prepared in a manner that will provide the ideal proliferation environment for given cells. For this reason, synthetic media can be divided in to four major categories. Serum containing media - In these types of media, serum fetal bovine serum is used as a carrier for nutrients and growth factors among others that tend to be water insoluble.
Serum-free media - These types of media is typically produced for the purposes of supporting single cell type of culture. As such, it provides specified nutrients and other factors required by the cell type. In this media, serum is absent because it present some disadvantages and can result in misinterpretation of immunological results. Chemically defined media - Like the name suggests, this type of media is composed of contamination- free pure organic and inorganic ingredients.
Protein-free media - Protein- free media are typically lacking of any type of protein. It is largely used to promote superior growth of the cells as well as protein expression in addition to facilitating for the purification of any expressed product. Some of the major components of cell culture media include;. Cell culture media are used for the proliferation of cells, which can then be identified and studied. As such, it can be used for various purposes including for education, diagnosis and treatment of a disease among others.
In culture methods, cell suspension refers to a type of culture where cells are suspended in a liquid medium. To obtain single cells, a friable callus small tissue that falls apart easily is put in agitated liquid medium agitation allows for gaseous exchange unlike solid medium , breaking it up.
This allows for single cells to be released, which are then transferred to another fresh medium. Cell suspension cultures have a big advantage over the stationary ones given that it allows for the cells to be uniformly bathed. Moreover, given that the medium tends to be agitated, it allows for aeration of the medium, providing gases to the cells. Given that the medium is a suspension, it also becomes easy to manipulate the contents of the culture.
Like any other culture, suspension cell culture has to be under controlled conditions, proving the cells with an ideal environment to proliferate. Once they reach about 80 percent confluence, it is time to subculture in order to ensure continued proper growth.
In some cases, the cells in suspension may adhere on to the plastic surface of the culture flask or even form clumps. In such cases, a pipette can be used to pick these cells and expel them on to the surface of the flask and therefore away from the plastic surface. This helps obtain single cells given that they are no adhere on to the plastic surface. Red Blood Cell Suspension. Counting the number of cells in a suspension is a process that involves the use of a stain.
For instance, when trypan blue is used, it penetrates the cell membrane of the dead cells, but not the living cells. The cells are then gently expelled into a haemocytometer contains the counting chamber under the cover slip and observed under a microscope.
Cells are then counted within a given number of squares for calculations. This method is largely preferred due to the fact that it allows for cells to be suspended in a solution rather than being held in a solid media.
Here, therefore, it becomes easier to manipulate the contents thereby preventing them from forming clusters. With cell suspensions, it is also easier to observe single cells under the microscope.
In this case, it becomes possible to not only study the structure of the cells, but also get to observe how well they have differentiated; viewing dead and living cells under the microscope.
Cell culture protocols are meant to ensure that culture procedures are carried out to the required standards. This is not only meant to prevent the contamination of the cells, but to also ensure that the researchers themselves are protected from any form of contamination. Moreover, the nature of the work is expected to conform to the appropriate ethical guidelines.
Therefore, before anything else, it is essential to ensure that the entire procedure conforms with both medical-ethical and animal- experiment guidelines. This is because going against such legislation and guidelines can result in heavy penalties and even shutting down of the laboratory. Before any work starts, carry out the following procedure;. Although there are a wide range of culture media for cells, it is important to keep in mind that cell cultures, and particularly primary cell cultures are easily prone to contamination in addition to the risk of containing undetected viruses.
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.
Protocols for cell culture preparation. 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 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 is 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 is 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. On the other hand, 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. In order to enhance the success of cultures, it is 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. Check out MicroscopeMaster's picks for Microscope Accessories. 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. Amazon and the Amazon logo are trademarks of Amazon. Scientific understanding changes over time. MicroscopeMaster is not liable for your results or any personal issues resulting from performing the experiment.
Introduction to Cell Culture
Note that while the basics of cell culture experiments share certain similarities, cell culture conditions vary widely for each cell type. Deviating from the culture. Cell culture involves the distribution of cells in an artificial environment (in vitro) composed of necessary nutrients, ideal temperature, gases, pH and humidity for . Cell culture has a diverse range of uses as cultured cells are used by cell biologists, biomaterials scientists, clinicians and regulatory authorities, among others.