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Exosome extraction from supernatant using Functional nanomagnetic fine particles

Research Project

Project/Area Number 15K15502
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Digestive surgery
Research InstitutionKagoshima 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
Project Status Completed (Fiscal Year 2017)
Budget Amount *help
¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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.

Report

(4 results)
  • 2017 Annual Research Report   Final Research Report ( PDF )
  • 2016 Research-status Report
  • 2015 Research-status Report

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Published: 2015-04-16   Modified: 2019-03-29  

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