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How do transfected stable cell lines advance biomedical research?



The important role of cell culture models for research progress

Experimental models should reflect the complex regulation of biological processes. However, they should also offer reproducible, standardized experimental conditions and reasonable timelines. Cell-based models provide a balance between these often conflicting requirements and have been established as key models to elucidate gene and cell functions. Moreover, cell-based models are indispensable for therapeutic target identification and high-throughput drug candidate screening.


How do primary cells and immortalized cell lines facilitate biomedical research?

Both primary cells and immortalized cell lines have been widely used in life science research and have advantages and limitations. Primary cells are isolated from living organisms and may reflect the investigated biological processes more closely. However, immortalized cell lines also offer important advantages, such as cost effectiveness, ease of use, homogeneity of the cellular population, and no ethical challenges.


Cell-based models as tools for pharmacological and toxicological research

Cell-based models, including both primary cells and cell lines, have found extensive applications in the fields of drug screening and toxicology as they enable efficient, high-throughput analysis. Cell-based models have played an essential role in the identification of numerous drug candidates that have subsequently progressed to evaluation in in vivo models. However, due to the high rate of failure of drug candidates during later stages of the drug development process, optimized cell-based models are needed.


How do molecular biological techniques improve cell-based experimental models?

Molecular biological manipulations can regulate gene expression through gene overexpression or knockdown and can play an important role in the elucidation of gene function. Moreover, transfection of reporter vectors can offer insight into the function of downstream cellular signaling pathways. Frequently, transient transfections are performed to achieve these goals. However, they result in gene expression changes with limited duration, rendering the experimentation process burdensome and time-consuming.


What are the advantages of stable gene expression- and reporter cell lines?

Transfected stable cell lines offer a solution for several of the challenges posed by transient transfections. To establish stable gene expression or reporter cell lines, the gene or reporter expression cassettes should be integrated into the cell genome using lentivirus or plasmid vectors. Thus, permanent gene integration is achieved, and no repeated transfections are needed. However, the development of stable cell lines is time-consuming, associated with high costs, and requires extensive professional expertise. Commercially available gene expression and reporter transfected stable cell lines enable researchers to focus on their scientific questions without wasting time and financial resources on stable cell line development.


Gene Expression Stable Cell Lines Expression and purification of eukaryotic proteins are necessary to elucidate their function and to enable their use in downstream applications but are technically challenging. Transfected stable cell lines have been developed as an experimental platform for the expression of such proteins. Subsequently, expressed recombinant proteins have been employed in various downstream applications, such as vaccine development. Moreover, cell lines stably expressing key genes may be used to elucidate their significance for various cell functions.


Reporter Stable Cell Lines Reporter cell assays have been established as valuable tools to identify ligands for cell signaling pathways and transcription factors as well as to screen for receptor agonists and antagonists.These cell lines have already contributed to major advances in the elucidation of transcription factor function or interactions. Moreover, they play an important role in the high-throughput preclinical screening of drug candidates.


Stable gene expression and reporter cell lines as valuable tools to elucidate cell functions

Transfected stable cell lines are indispensable tools for biomedical research. Even though the process to establish them is time-consuming and requires extensive expertise, stable cell lines provide valuable insight into molecular cell mechanisms and functions and may represent an efficient screening platform for drug candidates. Moreover, their maintenance is relatively simple and straightforward, and their use can be scaled up quickly, if the need arises.


Resources


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4. Chaudhary S, Pak JE, Gruswitz F, Sharma V, Stroud RM. Overexpressing human membrane proteins in stably transfected and clonal human embryonic kidney 293S cells. Nat Protoc. 2012;7(3):453–466. doi: 10.1038/nprot.2011.453.

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