The detection assay of activated platelets in human blood by flow cytometry

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K16944
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 52010:General internal medicine-related
• Research Institution: Hokkaido University (2020-2021); The University of Tokyo (2019)
• Principal Investigator: Yasumoto Atsushi 北海道大学, 大学病院, 臨床検査管理医師 (90769887)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 活性化血小板 / フローサイトメトリー

Outline of Final Research Achievements

(1) Platelet aggregates were formed by stimulating citrate blood with various platelet agonists, and measured with an optical / fluorescence microscope, a flow cytometer, and an Optofluidic time-stretch microscope. Density-gradient method and cell fixation enabled platelet aggregates to be detected with high sensitivity.
(2) Platelet aggregates showed CD61 + CD62P + PAC-1 + by a flow cytometer. Staining of CD45 is also important to evaluate the association between leukocytes and platelets. The results of this research were published in eLife (Zhou Y, Yasumoto A, et al. eLife. 2020; 9: e52938).
(3) We established a functional assay, which quantitatively evaluates the degree of platelet activation by measuring released microparticles.

Free Research Field

臨床検査学

Academic Significance and Societal Importance of the Research Achievements

脳梗塞や心筋梗塞は日本での死亡原因の大きな要因であり、その克服は重要課題である。それら血栓症の原因のひとつは動脈硬化であり、活性化血小板が関与する。動脈硬化が進行すると活性化血小板は増加すると考えられるが、その測定法はいまだに確立していない。本研究では生体内の活性化血小板をOptofluidic time-stretch顕微鏡という新しい技術を用いて検出して、臨床現場で使えるものを作り上げた。これが実臨床で用いられた場合、脳梗塞や心筋梗塞の発症リスクを早期に検出でき、予防が可能になるかもしれない。