High-throughput PBPK and microdosimetry: cell-level exposures in a virtual tissue context

Neurospheres are three dimensional (3D) cell clusters consisting of neural progenitor cells (NPCs). NPCs provide the cellular basis for the developing brain and persist in the hippocampus up to old age. there, they are responsible for adult learning and memory. Safety and efficacy testing with 3D neurospheres by employing high content image analyses (HCA) require advanced scanning and evaluation processes due to defocusing of the sphere core and sphere-specific endpoints. therefore, for this mixed neuron/glia culture we developed algorithms for radial migration, quantification of neuronal differentiation, neurite outgrowth and distance-dependent density distributions of neurons. For quantification of neuronal differentiation the new algorithm reaches an average detection power (DP) of 80-85% (versus manual evaluation) and a false positive rate (FP) of 10-15% improving results of the commercially available “Neuronal Profiling” bio-application (Thermo Scientific; DP: 50%, FP: 40%). This is due to the application domain of the “Neuronal Profiling”, which was initially designed for pure neuronal cultures. In conclusion, HCA of neurospheres is a promising technique for medium throughput screening to be used in safety and efficacy testing in the future.