Selection of cell types and assay validation for CardioXpress
10 Jan 2014

One of the biggest unmet needs in cardiac safety pharmacology is accurate high throughput prediction of adverse effects at an early stage in drug discovery. The sooner a potentially adverse compound effect is detected the sooner an adequate risk management strategy can be developed. Although several tests on isolated animal cardiac tissues including Purkinje fibers, papillary muscles and ventricular trabeculae are available, none is scalable enough for routine use in early drug discovery. Moreover the species barrier to humans makes that none of the animal based cardiac tissue assays is versatile and predictive enough to convince that a drug is deemed safe.

Human pluripotent stem cell derived cardiomyocyte assay technology has the potential to address this unmet need. A crucial part in assay development is the selection of the appropriate cardiomyocytes that closely resemble human adult cardiomyocyte biology. The major challenge in the field is to overcome maturity problems and to create these cells with minimal batch-to-batch variation. Fully functional cells do express the appropriate contractile proteins, the right ion channels and have a fully functional contraction-excitation coupling.

Pluriomics buildings in Leiden, the Netherlands

Pluriomics creates such a solution through the use of optimized chemically defined in every step of the manufacturing process, which ensures a reproducible mature phenotype with excellent pharmacological properties. Pluriomics cardiomyocytes are produced in large batches, conveniently cryopreserved and quality controlled for key electrophysiological parameters.

Importantly the use of optimized media allowed Pluriomics to optimize cardiomyocyte culture conditions for long-term stability using minimal medium changes. The lack of serum, which is a potent inducer of cardiac hypertrophy, is a major step forward. Finally, both acute and long-term assays can be performed in culture medium with a greatly reduced risk of skewed results due to unwanted plasma protein binding.

Human stem cell derived cardiomyocyte by Pluriomics. Immunolabelling of alpha actin (red) and troponin (green)

This knowledge is now applied to create a fully fine-tuned integrated Multi-electrode assay solution. To achieve this goal Pluriomics collaborates with Multi Channel Systems for multi-electrode technology in multi-well plates and NOTOCORD® for advanced automated field potential data analyses.

The project recently triggered an important milestone. Pluriomics has finalized their cardiomyocyte manufacturing, cryopreservation and culture procedures and is currently working on further assay development & validation using a panel of reference compounds and controls.

In conclusion Pluriomics managed to improve cardiomyocyte culture conditions considerably. Pluriomics and CardioXpress are making important steps towards an integrated beta prototype. Ultimately we believe that we will empower researchers to perform drug discovery research and cardiac safety pharmacology in parallel. The outcomes of each process can inform the decision making in the other, ultimately leading to more effective drug development.
 

CardioXpress received the Eurostars label. The Eurostars Programme is powered by EUREKA and the European Community.

     

Contact Stefan Braam directly.

 

Keywords

  • Human pluripotent stem cells
  • Cell culture
  • Serum free
  • Drug discovery