Cell Line Characterization. Biological & Biomedical Sciences Essay

Cell Line Characterization
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The field of biological and biomedical sciences is quite diverse and encompasses various aspects including cell line characterization. It is a critical concept that involves the control of and manufacturing of biological products. Cell line characterization is primarily inclined towards confirming not only the identity and purity but also the suitability of cell substrates required for manufacturing use. Similarly, research and development initiatives concerning cell lines warrant for the need to have precise knowledge about the purity and species or origin. In this regard, it is of utmost necessity to conduct periodic and regular monitoring of cultured cell lines to mitigate possible contamination. The monitoring initiative also helps in the determination of cell line authentication and identity. Cell line authentication is essential because it prevents the inadvertent contamination of cell lines in the course of regular cell culturing. During the drug development process, it is extremely important to authenticate both the identity and characteristics of a cell line to monitor the effects of new therapeutics and to ensure the maintenance of the parent cell. Scientists and biomedical experts note that failure to monitor cultured cell lines often results in the contamination of inter and intraspecies cell lines. Consequently, these contaminations result in mistaken and false conclusions. Experts further share that there are various strategies used in testing cell lines in efforts to characterize them. These strategies are based on different factors including origin of the cell lines and cultivation history. Additionally, cell line characterization must take into account several regulatory requirements such as product type, the type of product being manufactured using the cell lines and geographical region where the commercial licensure will be obtained. The essay seeks to explain in-depth the association of characterization of cell lines in process validation studies with focus on origin of cell lines, phenotyping, antibiotic resistance, identity and stability monitoring, testing for adventurous agents, retroviruses, retroviral activity markers and tumorgenecity.
Understanding Cell Line Characterization and Authentication
Biomedics and medical scientists are continuously involved in cell culturing efforts to address various health care concerns in society. According to Almeida, Cole and Plant (2016), “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 isolated from living tissue, they can subsequently be maintained under carefully controlled conditions” (p.e1002476). While the lifespan of most cells is genetically determined, there are some that have been exposed to optimal conditions transforming them into immortal cells that can reproduce indefinitely. The first stage of cells culture that sees the isolation of cells from animal or plant tissue is called primary culture (Ethier and Neve, 2015, p.102). The primary culture then becomes a cell line after it has been subcultured through transfer into an environment that supports its continued growth.
One primary issue that scientists face when working with cultured cells entails cell line cross-contamination. Research indicates that at least 20% of experiments pertaining to cell culture usually have cells that are either contaminated or misidentified (Ethier and Neve, 2015, p.103). Cross-contamination of cell lines has also been reported in drug screening studies. The issue is a concern for cell line repositories such as the American Type Culture Collection (ATCC) and the German Collection of Microorganisms and Cell Cultures. These institutions have in the past reported misidentified cell line submissions from researchers. In light of these problems, it has become important to conduct cell line characterization and authentication. Cell line characterization is a testing mechanism that confirms and determines the identity, purity and suitability of cell substrates used in manufacturing different drugs and medications. As such, it is a vital phase in controlling and ensuring only quality and standardized biological products are used in the manufacture of drugs that will be released to the market.
Similarly, cell line authentication is a crucial step that according to Ethier and Neve (2015), “assists to assure that inadvertent contamination of cell lines has not occurred during the regular cell culturing work” (p.104). It is highly recommended that cell line authentication is conducted at an early passage to determine the identity and to monitor the effects of new therapeutics during the drug development process. In this regard, cell line authentication is usually done repeatedly before freezing cell line stocks. It is done every two months during cell culturing and before any data generated from the cell lines is published. However, it is important to note that there are various regulatory requirements that need to be considered for cell line characterization. Some of these factors include product type and the geographical region where the commercial license will be obtained (Reid, 2009, p.14). For instance the US and members of the European Union have different regulatory requirements. It is also essential to consider the type of product that will manufactured using the cell lines since they can be used to manufacture antibodies and viral vaccines.
Origin of the Cell Line
As it was shared in the previous section, animal, human and plant cell cultures are significant tools in the scientific landscape. Reid (2009) notes that “Different variants of cell culture found application in modeling diseases, IVF technology, stem cell and cancer research, monoclonal antibody production, regenerative medicine and therapeutic protein production” (p.16). Based on the above statement, it is essential to note that all those scientific approaches would be impossible if not for the main milestones in cell cultures that have been witnessed over the years. Similarly, it has also been shared that a cell line is basically a permanently established cell culture that will grow and develop indefinitely so long it is placed in optimum conditions in the likes of fresh medium and space.
Scientists and biomedical experts have shared that a cell line is a product of cell culture. It is developed when the primary culture which is the first phase when cells are isolated from tissue are subcultured or passaged. The cell lines that are derived from the primary culture tend to have a limited life span. However, when these cells are placed in fresh growth medium, they are enhanced and given room to continue growing. These optimal conditions are characterized by vessels with essential nutrients such as amino acids, vitamins, hormones and regulation of the physio-chemical environment which includes osmotic pressure and temperature (Ethier and Neve, 2015, p.100). While some cells may require an artificial substrate, others can grow and develop in suspension cultures.
Additionally, in the production of biological products, it is essential to consider historical factors of the cell line. The first factor taken into account is the age, species and sex of the organism where the cells were harvested from. Secondly, in regards to human cell lines, it is important to determine the individual’s medical history. This initiative and the tests conducted will help to detect any adventitious agents on the donor. Thirdly, it is vital to understand the cell culture and eventual subculture of the cell line. On the same breadth, the methods used to isolate cells from the tissue should be considered. Last but not least, is also necessary to check previous identity tests and all the findings from adventitious agent testing. Furthermore, in the examination of the cell line, the general characteristics of the cell line should be considered. According to Reid (2009), “The growth pattern and morphological appearance of the cell line should be determined and should be stable from the master cell bank to the end-of-production cells” (p.17). For instance, all specific markers identified during cell line characterization should be examined for stability.
Antibiotic Resistance, Identity and Stability Monitoring
Antibiotics are biological and biopharmaceutical products that are manufactured to address various health care complications in human beings and animals. Particularly, they are used to prevent and treat bacterial infections. They are very strong medications that require prescribing and in some cases administering by medical professionals such as doctors and nurses. However, antibiotic resistance is a global threat not only to health but also food security and development in general. Antibiotic resistance affects individuals of all ages in various regions. It is estimated that at least 2 million Americans report antibiotic resistant infections and about 40000 out of that number die annually (McConnell, 2004, p.325). As a result of antibiotic resistance, a variety of infectious diseases such as pneumonia and tuberculosis are becoming impossible to treat because the drugs and medications used to treat them have become ineffective. It is also essential to note that antibiotic resistance not only results in higher health care costs and increased mortality but also makes patients to stay longer in hospital.
Scientists and medical experts have proposed that antibiotic resistance usually occurs from various factors. It has been shared that misuse of antibiotics in society is major contributor to resistance. These instances of misuse are noted when individuals ingest antibiotics for purposes other than what they were intended for. Similarly, it also occurs when patients fail to finish their antibiotic dosages. There is also another school of thought that is of the opinion that antibiotic resistance occurs naturally. According to McConnell (2004), “Antibiotic resistance is a consequence of evolution via natural selection. The antibiotic action is an environmental pressure; those bacteria which have a mutation allowing them to survive will live on to reproduce” (p.325). Based on the above statement, these mutated bacteria pass the trait to their offspring and in the long run, leads to the creation of a fully resistant generation. Similarly, overuse of broad-spectrum antibiotics also contributes in hastening resistance to methicillin.
Additionally, mishandling and mismanagement of bacteria, antibiotics during cell culture can also result in antibiotic resistance. Stollmeier and Nagler (2018) shares that “Antibiotics and/or antimycotic agents are added to cell culture media as a prophylactic to prevent contamination, as a cure once contamination is found, to induce the expression of recombinant proteins, or to maintain selective pressure on transfected DNA (p.10). Similarly, as it was shared in the previous section, cells are usually harvested from animal, plant or human tissues and placed in optimum conditions. However, there are cases when the solution used to keep the collected cells and ensure their continued growth and immortality is impure. Consequently, failure to realize the problem early enough will result in distorted results. In this case, it will lead to the creation of antibiotics that instead of addressing particular health care concerns, they are contributing in fostering resistance. Antibiotic resistant germs are known to multiply rapidly and in the process not only pass their resistance attributes to other germs but also kill good bacteria (Stollmeier and Nagler, 2018, p.19). Contrary to popular opinion, not all bacteria are harmful and there are some that help to protect the body from infection.
Based on the above facts, it is evident that many researchers often use antibiotics in their cell cultures to avoid bacterial contamination. While antibiotics are instrumental in that regard, reports indicate that they can have fuel the effects of contamination complications. In most cases, contamination of cell cultures usually occurs when fungi or bacteria is accidentally introduced into the mixture. These accidents can happen at any point during the handling and management of the cell cultures. Some of the common sources include aerosols, dust and mishandled reagents. There are three distinct outcomes and consequences of adding antibiotics in a cell culture. Firstly, it may result in resistance to antibiotics. This happens when one of the contaminating microorganisms is entirely resistant to the antibiotics used in the medium. Eventually, the contaminants grow and develop into immortal cells with serious health effects. The second consequence involves partial resistance to antibiotics. As a result of the...