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.
Also, because the sensor could track single cells in a 3D matrix, functions such as individual cell adhesion, spreading, and migration could be monitored. 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.
In the past several years, a huge variety of 3D cell culture systems have been created as experimental tools for diverse research purposes. There is no doubt that 3D culture systems hold great promise for applications in drug discovery, cancer cell biology, stem cell research, and many other cell-based analyses and devices, by bridging the traditional 2D monolayer cell culture to animal models. While the 3D cell culture models are currently widely studied in academia with a focus on creation of 3D systems with excellent biological relevance, there are still many hurdles that must be overcome before these systems can be widely accepted in industry.
Recent developments clearly indicate that the transition from 2D to 3D cell cultures for industry applications is promising, but the maturity of the technology and the cost are still the main concerns in making this transition possible. Much effort is still needed to assure reproducibility, high throughput analysis, compatible readout techniques, and automation in order to establish standardized and validated 3D cell culture models.
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Copyright , Mary Ann Liebert, Inc. This article has been cited by other articles in PMC. Abstract Three-dimensional 3D cell culture systems have gained increasing interest in drug discovery and tissue engineering due to their evident advantages in providing more physiologically relevant information and more predictive data for in vivo tests. Introduction Cell-based assays have been an important pillar of the drug discovery process to provide a simple, fast, and cost-effective tool to avoid large-scale and cost-intensive animal testing.
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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Once the cells have been obtained, they can either be cultured as explants culture, suspension or monolayer. That is, they are not from another culture process. Before the cells are cultured, they are first subjected to enzymatic treatment for dissociation. However, has to be for a minimal amount of time to avoid damaging or killing the cells.
Once single cells are obtained, they are then appropriately cultured in media to allow them to grow divide are reach the desired numbers. Initially, the culture tends to be heterogeneous in that it is composed of different types of cells obtained from the tissue.
Although this can be maintained through the in vitro process in a culture in a suitable media this would only be for a limited period of time. However, through the transformation process, the primary cells may be used for a long period of time, changing the culture over time.
These cells are refers to as continuous cell lines. However, primary cells are typically preferred over continuous cell lines because of the fact that they are more similar physiologically to in vivo cells cells from the living tissue. In addition, continuous cell lines may undergo certain changes phenotypic and genotypic changes which would result in discrepancies during analysis. As such, they cannot be used to determine what is happening to the in vivo cells. It is for this reason that primary cells are preferred.
Given that the primary cells significantly resemble the cells obtained from living tissue, they are important for research purposes in that they can be used to study their functions, metabolic regulations, cell physiology, development, defects and conditions affecting the tissue of interest.
In addition, they are used for such purposes as vaccine production, genetic engineering drug screening as well as toxicity testing and prenatal diagnosis among others. In cell culture techniques, cells or tissues are removed from a plant or an animal and introduced into a new, artificial environment that can support their proliferation survival and growth.
Some of the requirements of such an environment for the proliferation of the cells include;. Here, we shall focus on the medium cell culture media. Although there are different types of culture media for different types of cells they are typically composed of;. There are two major types of culture media. Natural media - Natural culture media is composed of biological fluids that are naturally occurring.
Although this type of media can be used for a range of cells, its biggest disadvantage is that it may lack the exact components required by given cells, which can greatly affect reproducibility. Artificial media - Also referred to as synthetic media, artificial media refers to the type of media that is produced by adding such nutrients as vitamins, gases oxygen and carbon dioxide and protein among others.
These organic and inorganic nutrients are added so as to meet the specific needs of given cells, and thus provide the ideal environment for their growth. 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.
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.