Caspase inhibitors increase the rate of recovery of neural stem/progenitor cells from post-mortem rat brains stored at room temperature |
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Authors: | Atsuko Hasegawa Chikako Yamada Miho Tani Shun-ichiro Hirano Yasuhito Tokumoto Jun Miyake |
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Affiliation: | 1Department of Tissue Engineering and Cell Therapy, Institute of Biomedical Research and Innovation, 1-5-4 Minatojima-Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan;2Research Institute of Cell Engineering, National Institute of Advanced Industrial Science and Technology, 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974, Japan;3Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan |
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Abstract: | 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 °C, 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 48 h at 15 °C, 24 h at 25 °C, in both condition, we were able to recover NSPCs from post-mortem rat brains. At 15 °C, 90% of neurosphere-forming activity was lost within 24 h. However, even after 24 h at 25 °C, 2% neurosphere-forming activity remained. After 6 h of death, there was very little difference between the rates of NSPC recovery at 4 °C and 25 °C. 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. |
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Keywords: | Neural stem cell Caspase inhibitor Caspase-8 Apoptosis Cell transplantation therapy |
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