Primary cells are isolated directly from human tissue and serve as a physiologically relevant pre-clinical model. They are a powerful tool for studying the effects of therapeutics on cell physiology and biochemistry (e.g. cell metabolism, ageing, cell health, etc…) and are an alternative to immortalized cell lines.
At Charnwood Discovery we have experience of utilizing a range of human primary cell types in a number of assay formats (freshly isolated PBMCs, Neutrophils, Whole Blood, Dermal Fibroblasts, HUVECs) and have HTA licensed premises for proper storage of human cells.
Increase the Relevance of your Assays
In Bioscience, we recognize that primary cells are more phenotypically relevant model in the drug discovery process. Primary cells better mimic in-vivo conditions; can improve relevance and reliability of cell-based assays and can serve as an alternative to animal models which are all key for the identification of drug candidates.
Utilizing these cells can improve the reliability of our assays to achieve more predictable data for in vitro ADME-Tox studies. There is now increased recognition of the importance of human primary in vitro cell based assays as a powerful tool in drug discovery.
As drug discovery programs develop, analysis of efficacy, pharmacokinetics, and pharmacodynamics become important for lead optimisation. Signalling pathways and protein levels of biomarkers can be examined in patient samples, xenograft models or samples from animal studies to gain understanding of therapeutic development in physiologically relevant model systems. These models are described as ‘ex vivo’, derived from the Latin “out of the living” and meaning to experiment on a tissue taken from an organism with minimal alteration of the physiological conditions.
At Charnwood Discovery, we have implemented the use of Simple Western™ technology in combination with sample homogenising technology such as the Bertin Technologies Precellys® Evolution homogeniser, to evaluate the therapeutic effect within a range of biological samples taken from living organisms.
Due to the conditions in which ex vivo samples are collected, their value and precious nature makes large quantities often difficult to obtain. The Simple Western technology requires as little as 0.6 µg of protein and is truly quantifiable, making it an ideal solution for analysis of precious samples.
Here is an example of our team using the Precellys® tissue homogeniser and Simple Western™ Jess immunoblotting to evaluate the levels of a phosphorylated protein target within kidney samples acquired from a diseased kidney mouse model. To achieve this, a fixed quantity of protein from each sample (as determined by BCA) is loaded and analysed with antibodies specific for the target of interest. .
Figure 1; Pseudoblot from the Simple Western™ showing replicates of sample groups.
Figure 2: Quantification of the phosphorylated target normalised to the total target derived from figure 1.
Ex vivo samples can also be analyzed with our DMPK team to evaluate compound levels from the same sample, which can provide a powerful diagnostic tool to determine the effect a therapeutic is having.
The combination of the Simple Western™ technology with improved ex vivo sample preparation (via the Precellys® Homogeniser) means that throughput for therapeutic development is greatly increased when compared to traditional methods, allowing for fast and reliable therapeutic development.
Find out how Charnwood Discovery can support your drug discovery program.