| 研究課題/領域番号 |
22K17135
|
|---|
| 研究機関 | 新潟大学 |
|---|
研究代表者 |
Thant Lay 新潟大学, 医歯学総合病院, 特任助教 (10941867)
|
|---|
| 研究期間 (年度) |
2022-04-01 – 2024-03-31
|
| キーワード | Extracellular vesicles / ROCK inhibitor / Bone regeneration |
|---|
| 研究実績の概要 |
MagCapture Exosome Isolation Kit was used to prepare Control extracellular vesicles (Control-EVs) and ROCK inhibitor-treated extracellular vesicles (ROCKin'-EVs) from the culture medium of BMSCs. Mass spectrometry was performed to elucidate the changes in the proteomic composition. We detected there were changes ROCKin'-EVs protein composition. We identified that a number of Rab GTPase (known as the key regulators of intercellular vesicle transport) were upregulated in ROCKin'-EVs.
We isolated the Control-EVs and ROCKin'-EVs from the culture medium of BMSCs again using total exosome isolation aiming to collect large amount of EVs. Western blotting of Control-EVs and ROCKin'-EVs was performed. The quantity of isolated EVs was inconsistent and necessary to be normalized.
|
| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
In order to collect large amount of EVs, total exosome isolation (Invitrogen) was used. However, normalizing the amount of EVs by this protocol is challenging. In order to overcome this, we will isolate and repeat the investigation of mineralizing activity of ROCKin'-EVs on osteoblasts using MagCapture Exosome Isolation Kit.
|
| 今後の研究の推進方策 |
We will investigate the mineralizing activity of ROCKin'-EVs on osteoblasts using MagCapture Exosome Isolation Kit. To evaluate the mineralizing activity, ALP staining, Alizarin Red S staining will be performed. To evaluate the expression levels of osteogenic genes (RUNX2, ALP), western blotting will be performed. Examination of bone regeneration ability of ROCKin'-EVs in rat model will be performed after reconfirming the In Vivo mineralization activity.
|
| 次年度使用額が生じた理由 |
We plan to continue both in vitro and in vivo research during next academic year (2023-2024). We will use the finance to purchase the necessary reagents, equipment for this purpose.
|
