Title:
METHOD FOR DETECTING NEUTROPHIL CHEMOTAXIS
Document Type and Number:
WIPO Patent Application WO/2022/061570
Kind Code:
A1
Abstract:
A method for detecting neutrophil chemotaxis, comprising: collecting a blood sample into an anticoagulant tube, adding glucose, uniformly mixing, and standing; taking the supernatant into a centrifuge tube, and centrifuging same; adding 1× calcium and magnesium ion-free Hanks balanced salt solution into the centrifuged blood sample to re-suspend cell aggregates at the bottom; uniformly blowing and dispersing same, and slowly adding a polysucrose solution from the bottom of the centrifuge tube to perform density gradient centrifugation treatment; the solution after density gradient centrifugation being divided into three layers, pipetting the clear liquid at the upper layer and the PBMC layer at the middle layer to obtain cell aggregates at the bottom layer; performing lysis treatment on erythrocytes in the cell aggregates by using sterile water, performing gentle blowing and suction, then successively adding 2× calcium and magnesium ion-free Hanks balanced salt solution and 1× calcium and magnesium ion-free Hanks balanced salt solution, uniformly mixing same, and performing centrifugation treatment again to obtain neutrophils; and counting the neutrophils, and pipetting the neutrophil suspension into a chemotaxis model for chemotaxis. The method can improve the speed of neutrophil extraction and shorten the detection time.
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Inventors:
SUN BINGWEI (CN)
YANG YUNXI (CN)
LIU LU (CN)
YANG YUNXI (CN)
LIU LU (CN)
Application Number:
PCT/CN2020/117066
Publication Date:
March 31, 2022
Filing Date:
September 23, 2020
Export Citation:
Assignee:
SUZHOU MUNICIPAL HOSPITAL (CN)
International Classes:
C12N5/071
Domestic Patent References:
| WO2007064379A2 | 2007-06-07 | |||
| WO2015132774A1 | 2015-09-11 | |||
| WO2008109816A1 | 2008-09-12 |
Foreign References:
| CN109971819A | 2019-07-05 | |||
| CN104513807A | 2015-04-15 | |||
| CN108676776A | 2018-10-19 | |||
| CN102321582A | 2012-01-18 | |||
| CN103160465A | 2013-06-19 |
Other References:
PETER M. DODEK, MASHIRO OHGAMI, DIANE K. MINSHALL , ALAN R. BURNS: "One-step isolation of neutrophils using an elutriator", IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - ANIMAL, vol. 27A, no. 3, 31 March 1991 (1991-03-31), US , pages 211 - 214, XP009535446, ISSN: 0883-8364, DOI: 10.1007/BF02630918
R. M. LOWENTHAL, D. S. PARK: "The Use of Dextran as an Adjunct to Granulocyte Collection with the Continuous-Flow Blood Cell Separator", TRANSFUSION, vol. 15, no. 1, 1 January 1975 (1975-01-01), US , pages 23 - 27, XP009118497, ISSN: 0041-1132, DOI: 10.1046/j.1537-2995.1975.15175103505.x
L. GLASSER , R. L. FIEDERLEIN, D. W. HUESTIS: "Granulocyte concentrates. Glucose concentrations and glucose utilization during storage at 22℃", TRANSFUSION, vol. 25, no. 1, 1 January 1985 (1985-01-01), pages 68 - 69, XP055916597, ISSN: 0041-1132, DOI: 10.1046/j.1537-2995.1985.25185116508.x
R. M. LOWENTHAL, D. S. PARK: "The Use of Dextran as an Adjunct to Granulocyte Collection with the Continuous-Flow Blood Cell Separator", TRANSFUSION, vol. 15, no. 1, 1 January 1975 (1975-01-01), US , pages 23 - 27, XP009118497, ISSN: 0041-1132, DOI: 10.1046/j.1537-2995.1975.15175103505.x
L. GLASSER , R. L. FIEDERLEIN, D. W. HUESTIS: "Granulocyte concentrates. Glucose concentrations and glucose utilization during storage at 22℃", TRANSFUSION, vol. 25, no. 1, 1 January 1985 (1985-01-01), pages 68 - 69, XP055916597, ISSN: 0041-1132, DOI: 10.1046/j.1537-2995.1985.25185116508.x
Attorney, Agent or Firm:
SUZHOU JINHE INTELLECTUAL PROPERTY AGENCY (SPECIAL GENERAL PARTNERSHIP) (CN)
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