Caspase inhibitors increase the rate of recovery of neural stem/progenitor cells from post-mortem rat brains stored at room temperature(CELL AND TISSUE ENGINEERING)

概要

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To match the demand of regenerative medicine for nerve system, collection of stem cells from the post-mortem body is one of the most practical ways. In this study, the storage condition of the post-mortem body was examined. We prepared neural stem/progenitor cells (NSPCs) from post-mortem rat brains stored at different temperatures. When brains were stored at 4℃, for one week, we were able to obtain neurospheres (a spheroid body containing NSPCs) by stimulation of cells with epidermal growth factor (EGF). Incremental increases in storage temperature decreased the rate of appearance of neurospheres. Within 48h at 15℃, 24h at 25℃, in both condition, we were able to recover NSPCs from post-mortem rat brains. At 15℃, 90% of neurosphere-forming activity was lost within 24h. However, even after 24h at 25℃, 2% neurosphere-forming activity remained. After 6h of death, there was very little difference between the rates of NSPC recovery at 4℃ and 25℃. Addition of caspase inhibitors to both the rat brain storage solution and the NSPC culture medium increased the rate of neurosphere-forming activity. In particular, an inhibitor of caspase-8 activity increased the NSPC recovery rate approximately three-fold, with no accompanying detrimental effects on neural differentiation in vitro.