論文 - 詳細
| RRC ID | 64333 |
|---|---|
| 著者 | Chua P, Lim WK. |
| タイトル | Optimisation of a PC12 cell-based in vitro stroke model for screening neuroprotective agents. |
| ジャーナル | Sci Rep |
| 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 neuronal cells as an in vitro stroke model. Nerve growth factor (NGF)-differentiated PC12 pheochromocytoma cells are frequently used. However, investigators often find their clonal variant undifferentiable and are uncertain of optimal culture conditions. Hence we studied 3 commonly used PC12 variants: PC12 Adh, PC12 from Riken Cell Bank (PC12 Riken) and Neuroscreen-1 (NS-1) cells. We found DMEM the optimal media for PC12 Riken and NS-1 cells. Using a novel serum-free media approach, we identified collagen IV as the preferred adhesive substrate for both cell lines. We found PC12 Adh cells cannot attach without serum and is unable to differentiate using NGF. NS-1 cells differentiated to a maximal 72.7 ± 5.2% %, with substantial basal differentiation. We optimised differentiated NS-1 cells for an in vitro stroke model using 3 h of OGD resulting in ~ 70% viable cells. We screened 5 reported neuroprotectants and provide the first report that serotonin is antiapoptotic in a stroke model and the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) is neuroprotective in PC12 cells. Thus we demonstrate the optimisation and validation for a PC12 cell-based in vitro stroke model. |
| 巻・号 | 11(1) |
| ページ | 8096 |
| 公開日 | 2021-4-14 |
| DOI | 10.1038/s41598-021-87431-4 |
| PII | 10.1038/s41598-021-87431-4 |
| PMID | 33854099 |
| PMC | PMC8046774 |
| MeSH | 8-Hydroxy-2-(di-n-propylamino)tetralin / pharmacology Animals Apoptosis / drug effects Cell Differentiation / drug effects* Cell Hypoxia Cell Survival / drug effects Culture Media, Serum-Free / chemistry Models, Biological* Neuroprotective Agents / pharmacology* Neuroprotective Agents / therapeutic use PC12 Cells Rats Serotonin / pharmacology Serotonin 5-HT1 Receptor Agonists / pharmacology Stroke / drug therapy Stroke / pathology |
| IF | 3.998 |
| リソース情報 | |
| ヒト・動物細胞 | PC-12(RCB0009) |