Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents

Abstract Stroke causes death and disability globally but no neuroprotectant is approved for post-stroke neuronal injury. Neuroprotective compounds can be identified using oxygen glucose deprivation (OGD) of cells as an in vitro stroke model. Nerve growth factor (NGF)-differentiated PC12 pheochromocytoma are frequently used. However, investigators often find their clonal variant undifferentiable uncertain optimal culture conditions. Hence we studied 3 commonly used variants: Adh, from Riken Cell Bank (PC12 Riken) Neuroscreen-1 (NS-1) cells. We found DMEM the media NS-1 Using a novel serum-free approach, collagen IV preferred adhesive substrate both cell lines. Adh cannot attach without serum unable to differentiate NGF. differentiated maximal 72.7 ± 5.2% %, with substantial basal differentiation. optimised model h OGD resulting ~ 70% viable screened 5 reported neuroprotectants provide first report that serotonin antiapoptotic 5-HT 1A agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) neuroprotective Thus demonstrate optimisation validation cell-based