Physiological Relevance

Cellular models are a main component of drug discovery as they are used all along the drug value chain, from target identification to in vitro ADMETox. Ideally, cellular models would be organotypic and mimicking the in vivo conditions: normal and disease physiology, genetic polymorphism, multicellular and 3D organization, and all organ models would communicate (“human-on-a-chip”) through microfluidics or bioreactors.

Although complex tissue models reflect the in vivo situation more reliably than simpler 2D or 3D systems, the higher the complexity, the lower the throughput. Highly physiological models also raise other problems such as validation, reproducibility, image analysis, maintenance, as well as costs.

At CYTOO, we are trying to reconcile quality and quantity with a bottom up approach: use high-throughput compatible cell models and bring more physiology into them, while keeping their throughput.

Micropatterning to improve physiology, two ways

Micropatterning restitutes physical cues to the cells for a restored mature phenotype. We have demonstrated higher level of differentiation and maturation compared to conventional culture conditions with several micropatterned cellular models:

  • myotubes: primary human myoblasts were pushed towards myotube differentiation and unprecedented level of striation,
  • iPS-derived cardiomyocytes were micropatterned to achieve sarcomere alignment, cell-cell connections with higher level of connexin 43, and synchronized beating,
  • kidney proximal tubes demonstrated basal and apical polarity around a true lumen while expressing high levels of drug and peptide transporters (OAT1, OCT2 and megalin), and increased CYP3A4 activity,
  • HepG2 C3A derived bile canaliculi showed increased metabolic activity and reactive metabolite detection.

Another advantage of micropatterning is to enable the formation of functional 3D spheroids through culturing (and not centrifugation). The 500um spheroids displayed a necrotic center, and an intermediate layer of quiescent cells surrounded by a proliferating zone. This 3D spheroid model mimics the growth of tumors in vivo, and enables the assessment of drug activity on quiescent cells, a potential reservoir for metastasis, not possible while screening only on proliferating monolayers.

Let us bring more physiology to your cell-based assays! Contact us now.