Human induced pluripotent stem cells (hiPSC) are obtained by genetically reprogramming somatic cells from human tissues, after which these cells can be further differentiated virtually into any cell types. hiPSC are derived from normal adults or from diseased patients and carry protein mutations that are able to compromise the cell physiology.
As consequence, hiPSC-derived cells reveal a high potential for studying pathophysiological processes in basically all pathological areas including CNS, PNS, peripheral tissues (cardiac, hepatic, pulmonary, muscular, vessel, blood, etc.) as well as for studies on tissue regeneration or toxicology. In addition to the big advantage of their human origin, they possess characteristics similar to primary cells which are usually non-human and have variability due to the differences in animal genetics.
hiPSC-derived cell systems are widely used in drug discovery projects by pharma, biotechs, or CROs typically for secondary (phenotypic) but also in primary drug screening assays in HTS mode, and are potentially useful for translational studies (animal to human).
Selvita has developed an in vitro model of neurodegeneration using terminally differentiated neuron cells, a population of human glutamatergic neurons derived from hiPSC. Cryo-preserved iCell®GlutaNeurons (Fujifilm-Cellular Dynamics), following thawing and plating, developed neurite-like branched networks within 24 hours and remain viable and adherent for an extended period in culture of >14 days.
In the figures below are shown results of the iCell GlutaNeuron-based assay development and optimization of an excitotoxic model of neurodegeneration, useful for identification of potential neuroprotectants. The hiPSC-derived neurons were treated with glutamate in presence or absence of AMPA and NMDA receptors antagonists as tool compounds (negative controls) for assay validation:
Fig. A. iCell GlutaNeurons plated at 40k cells per well, were grown up to 14 days in vitro (DIV) in 96 wells plates, then used for experiments. To note the progressive development of a complex neuritic network during the culturing period. The protocol used for cell culture induces development of ≥90% pure population of human glutamatergic neurons expressing typical neuronal markers (not shown).
Fig. B. Optimization of conditions for the development of the iCell GlutaNeurons-based neurodegeneration assay (96 well plates). Differentiated neurons were challenged with glutamate in concentration response for 48 hours. Two readouts (ATP levels and LDH release) were used to determine the neurotoxic effects, with similar results.
Fig. C. Effects of AMPA and NMDA receptors antagonists on glutamate induced toxicity on 14DIV hiPSC-derived glutamatergic neurons.