Exosome extraction from supernatant using Functional nanomagnetic fine particles

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K15502
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Digestive surgery
• Research Institution: Kagoshima University
• Principal Investigator: MORI Shinichiro 鹿児島大学, 医歯学域附属病院, 助教 (00620519)
• Co-Investigator(Kenkyū-buntansha): 奥村 浩 鹿児島大学, 医学部・歯学部附属病院, 助教 (10398282) ; 前村 公成 鹿児島大学, 医学部・歯学部附属病院, 准教授 (30398292) ; 喜多 芳昭 鹿児島大学, 医学部・歯学部附属病院, 助教 (30570692) ; 馬場 研二 鹿児島大学, 附属病院, 医員 (30642615) ; 夏越 祥次 鹿児島大学, 医歯学域医学系, 教授 (70237577)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: エクソソーム / 大腸癌幹細胞 / EMT / miRNA / ナノ磁気ビーズ

Outline of Final Research Achievements

(Method)HCT116 cells washed with phosphate-buffered saline (PBS), and the culture medium replaced with RPMI medium without FBS. After incubation for 24 h, the supernatant collected and centrifuged at 2,000 g for 10 min at 4 °C. Five ml of supernatant containing 1 ml of the ExoQuick incubated 24 hour at at 4 °C and centrifuged at 1,500 g for 30 min. After centrifugation of the pellet at 1,500 g for 5 min at 4 °C, the supernatant removed again. The pellet added 1.5 ml of PBS for further SEM. Sample was placed on membrane grid for 10 min and then fixed using a solution of 2% of gluteraldehyde and 2% of paraformaldehyde in a 0.1 M of sodium cacodylate/HCl buffer pH 7.2. Specimen dried naturally and scanned for examination.
(Results and Discussion)We concentrated exosome derived from HCT116 using the ExoQuick and confirmed it via the scanning of electronic microscope. The ExoQuick was suitable for extraction of exosome without waste.

Free Research Field

消化器外科