Recently, a growing trend toward precision medicine has seen many researchers switch to using primary cells. Unlike immortalized cell lines that have acquired multiple genetic changes to allow their indefinite growth in culture, primary cells are isolated directly from tissue.
This means that primary cells maintain the true characteristics of the tissue source, providing researchers with a system that has greater translational relevance.
By working with primary cells, it’s possible to study the impact of donor specific traits such as age, genetics, or influences of medical conditions that may impact the functional phenotype of cells.
Furthermore, by comparing normal primary cells to those taken from an individual with a condition such as cancer, researchers can better understand disease development and progression, enabling identification of potential biomarkers, and more effective treatments.
Despite the many advantages that primary cells offer, working with them poses several major challenges. First, primary cells are difficult to obtain in large numbers and many do not divide, making further scale-up difficult.
Secondly, primary cells are less robust than immortalized cell lines, meaning they can tolerate only a narrow range of culture conditions, which need to be optimized, and require extremely careful handling to avoid damage by environmental fluctuations.
Some primary cell types don’t survive long in culture, while others rapidly differentiate or de-differentiate; this increases the pressure on researchers to generate data quickly.
Finally, primary cells are never 100% pure, making it essential to minimize the growth of contaminating cells that may potentially skew results. One way of addressing the issues inherent to working with primary cells is to consider using live cell imaging.
Methods that provide non-invasive, real-time evaluation of dynamic primary cell populations promise to improve experimental consistency by reducing cellular stress.
Using an automated system specifically designed to fit inside a conventional cell incubator, researchers can readily assess the health and phenotype of primary cells by monitoring morphology, growth properties and differentiation state within the native environment.
This allows data-driven decisions to be made while an experiment is still in progress.
A major advantage of using live cell imaging for primary cells is that it delivers a real-time visual assessment of cell morphology. This not only provides confidence in the cell type under study, but it also allows early identification of contaminating cells.
Correlating cell morphology to assay readouts lets researchers more easily explain any unexpected results. Live cell imaging can also be used to evaluate cell numbers and confluence.
This is important because over confluence can promote the growth of contaminating cells or may cause primary cells to differentiate.
Using live cell imaging to identify and maintain consistent cell doubling times, researchers can achieve more stable primary cell populations and more reliable assay performance.
Another benefit of live cell imaging is that it supports dynamic functional assessments of cell health. Cell health assays are essential to many research objectives.
These include evaluating specialty media; screening drug candidates; studying the cellular changes associated with disease; and identifying factors that affect processes such as immune cell activation or stem cell differentiation.
Being able to study cell health in real time offers a level of insight that simply cannot be achieved using end-point assays, which provide limited kinetic information.
Additionally, live cell imaging generates data that can be useful for cell line authentication and assists with quality assurance efforts by promoting consistency and reproducibility.
By increasing the applicability of established QC approaches to primary cell culture, live cell imaging is having a significant impact on experimental reproducibility. One area where live cell imaging has seen particular utility in recent years is primary neuronal cell culture.
This field is rapidly growing in importance for its relevance to regenerative medicine and the study of diseases such as Alzheimer’s and Parkinson’s. Because primary neurons are extremely delicate, monitoring them requires simple, cell sparing protocols.
Using live cell imaging to characterize neurite outgrowth, maturation, and the disruption of neurite networks, researchers can learn more about neurodegenerative conditions with the aim of developing effective therapies.
Live cell imaging of primary cells is also an effective approach to study virus infections.
Although primary cells are often used for this purpose, they frequently harbor latent viruses that may be reactivated when the primary cells are removed from the native environment.
Propagation of these latent viruses can cause various cytopathic effects that are easily confused with the pathogenesis of the target virus. These are exacerbated by adverse growth conditions that can further complicate disease diagnosis.
By performing live cell imaging with highly optimized growth media within the controlled conditions of an incubator, researchers offer primary cells a stable environment more representative of conditions within the host.
This not only helps to prevent latent viruses from multiplying, but it also avoids cell deterioration that can be mistaken for virus-driven effects.
Live cell imaging provides important kinetic and morphological information to help you get the most out of your primary cells. With this automated technology, cultures can be easily monitored over time without perturbation, providing greater consistency and scientific insight. Why not give it a go and see just how easy it can be?
More Information