Over the past few decades, cell-based assays have provided scientists with outstanding insights on various cellular processes taking place in the body. Primary cell-based assays have proved to be one of a kind in the field of assay development. Human primary cell assays are developed by extracting cells from blood, differentiated stem cells or bone marrow. Development of in vitro cell based assays for relevant diseases can provide us with ultimate solutions to foresee the translational success of the diseases. These cell -based assays play a significant role in drug discovery and in the treatment of genetic diseases, degenerative diseases and in extensive research based on biological, biochemistry and other metabolic processes going on within the cell.
Why in Vitro Assay Development of Human Primary Cells?
It is essential to develop assay based on human primary cells because it helps us in providing the details of a cell, for instance; cell viability, response of the cell to cytokine, proliferation of cells, biochemical functioning of the cells or to inspect whether there are any phenotypic changes in the cells. In vitro development of primary cells is an emerging field which is no less than a boon for regenerative medicines and revitalizing damaged tissues or/and organs of the body. These assays provide us the incentives for the complicated molecular activities and interactions taking place at the cellular level and offer relevant resources for stem cell therapy and discovery of novel drugs.
Successful Handling of Human Primary Cell-Based Assays
Primary cell-based assays have come out as an apparent and powerful inclusion to compliment other techniques which are appropriate for high-throughput screening. However, when handling them, one needs to be very cautious as they are tough to culture. There are several points which we should keep in mind while working with these assays:
- Assay Control
Assay control determines the proper functioning of the assays. It is important to use positive control, negative control and untreated primary cells as they provide us with reference point for the analysis of data.
- Optimization of Cell Density
Coherent density of cells in a sample helps in the determination of the range of the assay and provides appropriate sensitivity of the sample.
- No Contamination
Primary cells extracted from the blood, embryo or bone marrow can get contaminated by several pathogens which can lead to inconsistency in results. Even the tiniest contaminant has the capacity to destroy weeks of studies.
- Cell Apoptosis
Whilst working with cell-based assay, it is essential to under the process of cell apoptosis or cell death. In the process of apoptosis; the metabolic activities of the cell stop; cytoplasmic material is released in the surrounding and the cell dies. Since primary cells are not immortal, it is important to understand the physiology of the cell type being used to obtain perfect results.
- Environmental Factors
Maintaining an appropriate temperature is a vital step in the assay development of primary cells. Even the slightest variations in the environment can severely affect the cell-based assay. It is essential to maintain appropriate CO2, if the concentration of CO2 is very high in the incubator; it may lead to poor cell viability.
In conclusion, human primary cell-based assays are emerging as an effective tool for drug discoveries and for the treatment of several deadly diseases. No wonder these cell-based assays are the tipping point in scientific advancements. Although, they come up with some challenges which needs to be tackled precisely.