Aims: When cerebral cortical neurons are damaged due to stroke or trauma, the injured neurons typically do not recover, resulting in persistent sequelae such as motor paralysis. One potential therapeutic approach is the transplantation of neurons derived from pluripotent stem cells. While we have demonstrated that cerebral organoids can be differentiated from human ES cells, using human iPS cells is more desirable for clinical applications due to the possibility of allogeneic transplantation with HLA compatibility. Here, we will perform non-clinical trials to establish differentiation protocols from hiPSCs to cerebral cortical neurons and assess the safety and efficacy of these differentiated cells. Additionally, we will explore large-scale cell production for future industrialization.

Methods: Organoids were dissociated into single cells with Neuron Dissociation Solutions. Dissociated cells were re-suspended with HBSS supplemented with 10% (v/v) KSR and 10μM Y-27632 at 1,000 cells/µL density. Cell suspension was loaded onto a Chromium Next GEM Chip G (2000177 10X Genomics), targeting 3,000 cells for capture per well, and processed in the Chromium controller to obtain Gel Beads-in-Emulsion. Libraries were generated and RNA-seq was performed.

Targets: Organoids derived from induced pluripotent stem cells of a single healthy individual.