WO2002073190A1 - Procede de triage de particules au moyen de gravites specifiques differentes - Google Patents

Procede de triage de particules au moyen de gravites specifiques differentes Download PDF

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Publication number
WO2002073190A1
WO2002073190A1 PCT/JP2002/002317 JP0202317W WO02073190A1 WO 2002073190 A1 WO2002073190 A1 WO 2002073190A1 JP 0202317 W JP0202317 W JP 0202317W WO 02073190 A1 WO02073190 A1 WO 02073190A1
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WIPO (PCT)
Prior art keywords
specific gravity
centrifuge tube
fine particles
solid
solid inclusions
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PCT/JP2002/002317
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English (en)
Japanese (ja)
Inventor
Hajime Ogata
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Hajime Ogata
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Publication date
Application filed by Hajime Ogata filed Critical Hajime Ogata
Publication of WO2002073190A1 publication Critical patent/WO2002073190A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material

Definitions

  • the present invention relates to a method and an apparatus for separating a mixture of two or more types of fine particles having different specific gravities into two groups of light and heavy weights simply and with high accuracy.
  • the present invention relates to a method and apparatus suitable for removing erythrocytes from blood and separating and collecting nucleated cells such as leukocytes.
  • Conventional methods for removing erythrocytes from body fluids such as blood to separate and collect nucleated cells include (1) a buffy coat-coating method, (2) a red blood cell removal method by hemolysis, and (3) a hemagglutination sedimentation. Separation method, (4) Centrifugation method using density gradient of specific gravity solution, (5) Separation method of high specific gravity using partition wall fixed in centrifuge tube, (6) Floating solid inclusion for centrifugation Techniques such as a centrifugal separation method using a method are known.
  • This method is a method performed by the present inventor and already published as Japanese Patent Application Laid-Open No. Hei 9-155552, the purpose of which is to centrifuge together two or more types of particles having different specific gravities.
  • a solid inclusion that intervenes in the specific gravity band can move up and down in the centrifuge tube by centripetal force to separate particles into two groups of light and heavy, and has a concave or sloped upper surface whose size just fits the inner wall of the centrifuge tube. It is used.
  • Such a solid inclusion has a large contact area with the wall of the centrifuge tube, and a centrifugal force is applied obliquely outward to the solid inclusion against the inner wall of the centrifuge tube during centrifugation.
  • centrifugal force and specific gravity setting conditions for intervening between the erythrocyte layer and the nucleated cell layer after centrifugation are narrow, and further improvements are expected. It is rare.
  • the present inventor has found that a fine particle group containing two or more kinds of fine particles having different specific gravities and a solid inclusion having a specific specific gravity and a shape are centrifuged together, so that the method is more efficient and more efficient than the conventional method.
  • a method for separating fine particles with different specific gravities into two groups with light inclusions in Sakai.
  • a fine particle group containing two or more types of fine particles with different specific gravities has a specific gravity located in the middle zone of the specific gravity of the fine particle group, and can move up and down while inscribed in the centrifuge tube. Centrifuge with the solid inclusions whose upper part is more convex than the contacting part, and pass the tangent line between the inner wall of the centrifuge tube and the solid inclusions to move high specific gravity fine particles below the solid inclusions or low specific gravity.
  • a method for separating fine particles having different specific gravities wherein the fine particles are moved above the solid inclusions and at the same time, the solid inclusions are moved and interposed between the high and low density two particle groups,
  • two or more kinds of fine particles having different specific gravities to be separated are fine particles.
  • Particles having a diameter of about 0.01 to 200 / zm and containing two or more kinds of fine particles with different specific gravities regardless of the type and shape of the fine particles, especially fine particles are mixed in the liquid medium It is a fine particle group in a state.
  • a typical example is a body fluid, particularly blood containing cells having various specific gravities such as red blood cells and white blood cells.
  • Other examples include colloidal liquids containing colloidal particles having different specific gravities, liquids containing micelles, emulsions, suspensions, and the like.
  • it is particularly suitable for collecting nucleated cells by separating and removing red blood cells from a cell population in a body fluid.
  • the material, size and the like of the centrifuge tube used in the present invention are not particularly limited, as long as they are used in an ordinary centrifuge. That is, it is preferable that the bottom 6 has a shape such as a hemispherical shape, a semiellipsoidal shape, or a cone shape, and a cylindrical body following the bottom 6.
  • the material of the centrifuge tube should be a material that has a gap that allows the particles to move between the wall of the centrifuge tube and the solid inclusion inscribed when a centrifugal force is applied, or such a gap. What has elasticity enough to produce Examples of the hard material include glass and metal materials, and examples of the material having elasticity include synthetic resin and rubber.
  • the inner wall of the centrifuge tube may be coated with a lubricant such as paraffin so that solid inclusions can easily move up and down the centrifuge tube during centrifugation.
  • the solid inclusion used in the present invention is, when two or more types of fine particles having different specific gravities are dispersed in a liquid, insoluble in the liquid, and when a centrifugal force is applied, the fine particles are in contact with the centrifuge tube wall.
  • a material having a gap that can move between the solid inclusions inscribed therein and a material having flexibility enough to create such a gap, that is, an elastic body is preferable.
  • Examples of the material of the solid inclusion 2 include agar, gelatin, a silicone resin, a polystyrene resin, a synthetic resin such as an ABS resin and a polyurethane resin, ceramics such as glass, proteins, and polysaccharides.
  • a cavity may exist inside the solid inclusion, and the cavity may be filled with another substance.
  • One of the features of the present invention is that the shape of the solid inclusions present at the interface of the fine particles having different specific gravities by centrifugation becomes convex above the part inscribed in the centrifuge tube. It was molded as follows.
  • Specific shapes of the solid inclusion include those formed in a line with the inner wall of the centrifuge tube, such as a true sphere, an oval sphere, a foot pole, a raindrop, a spindle, and an abacus. Among them, particularly preferred is a true sphere or ellipsoid (the ratio of the major axis to the minor axis is preferably within 1: 2).
  • the specific gravity of the solid inclusions can be appropriately selected depending on the purpose of separating fine particles.
  • a substance having a specific gravity that falls between the specific gravities of two fine particles to be separated for example, silica, clay
  • the specific gravity can be adjusted arbitrarily by mixing the required amount of ceramic powder, metal powder, etc., making the interior hollow, and enclosing other substances in the cavity.
  • a spherical solid inclusion 2 is placed in a centrifugal tube 1 having a hemispherical bottom 6 and a cylindrical body 7 following the same so as to inscribe the centrifugal tube 1.
  • Blood 10 containing two kinds of microparticles having different specific gravities of red blood cells 3 (high-density microparticles) and nucleated cells 4 (low-density microparticles) is placed in this centrifuge tube 1 (Fig. 4), and the centrifuge tube is centrifuged. .
  • red blood cells 3 having a high specific gravity gather at the part 5 where the inclined surface of the solid inclusion and the inner wall of the centrifugal tube are asymptotic, and when the centrifugal force is further applied, the red blood cells 3
  • the solid inclusions 2 move downward through the solid inclusions 5 while being pushed up by the negative centrifugal force generated by the specific gravity difference from the red blood cells.
  • most of the red blood cells 3 move below the solid inclusions, and the solid inclusions 2 intervene at the interface 9 where the nucleated cells 4 and the red blood cells 3 are in contact (FIG. 6).
  • a mixture of two or more types of fine particles having different specific gravities for example, blood, may be arranged below the solid inclusion.
  • a mixture of two or more kinds of fine particles having different specific gravities is put into a centrifuge tube, and a solid inclusion formed thereon is introduced.
  • the shape of the solid inclusion is preferably spherical or elliptical because of its easy insertion.
  • the microparticles antagonize the narrow space 5 formed by the asymptotics between the surface 8 of the solid inclusion and the inner wall of the centrifuge tube while leaving a small specific gravity gradient. Even if there is a lot difference, the desired low specific gravity fine particles 4 can be stably separated in a state where other fine particles are not mixed.
  • the liquid phase containing the low-specific-gravity particles above the solid inclusions can be easily separated by tilting the centrifuge tube or by using a suction tool such as a pipette to separate the high and low groups of fine particles. it can.
  • a polystyrene sphere having a diameter of 10.6 Oram (specific gravity: 1.05) was molded and inserted into a glass tube centrifuge tube having an inner diameter of 10.75 mm and a length of 75 mm.
  • 4 ml of human peripheral blood (specific gravity: about 1.056) was poured into the centrifuge tube, and the mixture was centrifuged at 50 OG for 20 minutes in a centrifuge.
  • Erythrocytes (specific gravity: 1.0964) that settled on the inner tangent to the solid inclusions moved down the solid inclusions along the inner wall of the centrifuge tube, and simultaneously the solid inclusions were pushed upward. Eventually, a fraction containing serum and leukocytes remained above the solid inclusions, and red blood cells accumulated below.
  • a fine particle group containing two or more types of fine particles having different specific gravities can be converted into two groups with high and low specific gravities with high accuracy by simply performing a conventional centrifugal separation operation using a relatively simple apparatus. Can be fractionated.
  • this method is very suitable for removing red blood cells from blood and collecting leukocytes, which are nucleated cells.

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Biochemistry (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Centrifugal Separators (AREA)

Abstract

L'invention concerne un procédé permettant de trier facilement et précisément les groupes de particules renfermant deux ou plusieurs types de particules, au moyen de gravités spécifiques différentes, en deux groupes : un groupe à gravité spécifique élevée et un groupe à gravité spécifique basse. Ce procédé comprend l'étape consistant à séparer de manière centrifugée les groupes de particules conjointement avec des inclusions solides possédant une gravité spécifique située dans la zone intermédiaire de la gravité spécifique des groupes de particules, pouvant être déplacée de manière verticale pendant l'inscription dans un tube centrifuge, et comprenant une partie supérieure provenant d'une partie s'inscrivant dans le tube centrifuge formé dans la forme projetée, de manière à déplacer des particules à gravité spécifique élevée vers le côté inférieur des inclusions solides, via la ligne tangentielle de la paroi interne du tube centrifuge vers les inclusions solides ou à déplacer des particules à gravité spécifique basse vers le côté supérieur des inclusions solides et, en même temps, à déplacer les inclusions solides dans une position intermédiaire de deux groupes à gravité spécifique élevée et basse de particules, le procédé pouvant être mis en oeuvre de manière favorable pour séparer des leucocytes du sang.
PCT/JP2002/002317 2001-03-13 2002-03-12 Procede de triage de particules au moyen de gravites specifiques differentes WO2002073190A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2001-070651 2001-03-13
JP2001070651A JP2005098704A (ja) 2001-03-13 2001-03-13 比重の異なる微粒子の分別法

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WO2002073190A1 true WO2002073190A1 (fr) 2002-09-19

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Cited By (7)

* Cited by examiner, † Cited by third party
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US8394342B2 (en) 2008-07-21 2013-03-12 Becton, Dickinson And Company Density phase separation device
US8747781B2 (en) 2008-07-21 2014-06-10 Becton, Dickinson And Company Density phase separation device
US8794452B2 (en) 2009-05-15 2014-08-05 Becton, Dickinson And Company Density phase separation device
WO2015070273A1 (fr) * 2013-11-14 2015-05-21 Greiner Bio-One Gmbh Dispositif de réception, procédé de production de celui-ci et procédé de séparation d'un mélange
US9333445B2 (en) 2008-07-21 2016-05-10 Becton, Dickinson And Company Density phase separation device
US9682373B2 (en) 1999-12-03 2017-06-20 Becton, Dickinson And Company Device for separating components of a fluid sample
US9694359B2 (en) 2014-11-13 2017-07-04 Becton, Dickinson And Company Mechanical separator for a biological fluid

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US7832566B2 (en) 2002-05-24 2010-11-16 Biomet Biologics, Llc Method and apparatus for separating and concentrating a component from a multi-component material including macroparticles
US20030205538A1 (en) 2002-05-03 2003-11-06 Randel Dorian Methods and apparatus for isolating platelets from blood
US20060278588A1 (en) 2002-05-24 2006-12-14 Woodell-May Jennifer E Apparatus and method for separating and concentrating fluids containing multiple components
US7845499B2 (en) 2002-05-24 2010-12-07 Biomet Biologics, Llc Apparatus and method for separating and concentrating fluids containing multiple components
US8567609B2 (en) 2006-05-25 2013-10-29 Biomet Biologics, Llc Apparatus and method for separating and concentrating fluids containing multiple components
JP5479319B2 (ja) * 2007-04-12 2014-04-23 バイオメット・バイオロジックス・リミテッド・ライアビリティ・カンパニー ブイ式懸濁液分画システム
US8328024B2 (en) 2007-04-12 2012-12-11 Hanuman, Llc Buoy suspension fractionation system
EP2645107B1 (fr) * 2007-06-08 2018-02-28 Quest Diagnostics Investments Incorporated Procédé de purification des lipoprotéines approprié pour l'analyse par mobilité différentielle de particules chargées
EP2259774B1 (fr) 2008-02-27 2012-12-12 Biomet Biologics, LLC Procédés et compositions pour administrer un antagoniste de récepteur d interleukine-1antagonist
EP2254991B1 (fr) 2008-02-29 2018-08-22 Biomet Manufacturing, LLC Système et procédé pour la séparation d'une matière
US8313954B2 (en) 2009-04-03 2012-11-20 Biomet Biologics, Llc All-in-one means of separating blood components
US9011800B2 (en) 2009-07-16 2015-04-21 Biomet Biologics, Llc Method and apparatus for separating biological materials
US8591391B2 (en) 2010-04-12 2013-11-26 Biomet Biologics, Llc Method and apparatus for separating a material
JP6057251B2 (ja) 2011-11-11 2017-01-11 国立研究開発法人産業技術総合研究所 粒子分別装置および粒子分別方法
US9642956B2 (en) 2012-08-27 2017-05-09 Biomet Biologics, Llc Apparatus and method for separating and concentrating fluids containing multiple components
US20140271589A1 (en) 2013-03-15 2014-09-18 Biomet Biologics, Llc Treatment of collagen defects using protein solutions
US9895418B2 (en) 2013-03-15 2018-02-20 Biomet Biologics, Llc Treatment of peripheral vascular disease using protein solutions
US10208095B2 (en) 2013-03-15 2019-02-19 Biomet Manufacturing, Llc Methods for making cytokine compositions from tissues using non-centrifugal methods
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000237279A (ja) * 1998-12-05 2000-09-05 Becton Dickinson & Co 流体サンプルの成分分離用アセンブリ

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000237279A (ja) * 1998-12-05 2000-09-05 Becton Dickinson & Co 流体サンプルの成分分離用アセンブリ

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US9682373B2 (en) 1999-12-03 2017-06-20 Becton, Dickinson And Company Device for separating components of a fluid sample
US9714890B2 (en) 2008-07-21 2017-07-25 Becton, Dickinson And Company Density phase separation device
US9452427B2 (en) 2008-07-21 2016-09-27 Becton, Dickinson And Company Density phase separation device
US9700886B2 (en) 2008-07-21 2017-07-11 Becton, Dickinson And Company Density phase separation device
US8394342B2 (en) 2008-07-21 2013-03-12 Becton, Dickinson And Company Density phase separation device
US10350591B2 (en) 2008-07-21 2019-07-16 Becton, Dickinson And Company Density phase separation device
US9333445B2 (en) 2008-07-21 2016-05-10 Becton, Dickinson And Company Density phase separation device
US9339741B2 (en) 2008-07-21 2016-05-17 Becton, Dickinson And Company Density phase separation device
US9933344B2 (en) 2008-07-21 2018-04-03 Becton, Dickinson And Company Density phase separation device
US8747781B2 (en) 2008-07-21 2014-06-10 Becton, Dickinson And Company Density phase separation device
US9364828B2 (en) 2009-05-15 2016-06-14 Becton, Dickinson And Company Density phase separation device
US9731290B2 (en) 2009-05-15 2017-08-15 Becton, Dickinson And Company Density phase separation device
US8998000B2 (en) 2009-05-15 2015-04-07 Becton, Dickinson And Company Density phase separation device
US11786895B2 (en) 2009-05-15 2023-10-17 Becton, Dickinson And Company Density phase separation device
US10807088B2 (en) 2009-05-15 2020-10-20 Becton, Dickinson And Company Density phase separation device
US9919307B2 (en) 2009-05-15 2018-03-20 Becton, Dickinson And Company Density phase separation device
US9802189B2 (en) 2009-05-15 2017-10-31 Becton, Dickinson And Company Density phase separation device
US9919309B2 (en) 2009-05-15 2018-03-20 Becton, Dickinson And Company Density phase separation device
US9919308B2 (en) 2009-05-15 2018-03-20 Becton, Dickinson And Company Density phase separation device
US11351535B2 (en) 2009-05-15 2022-06-07 Becton, Dickinson And Company Density phase separation device
US8794452B2 (en) 2009-05-15 2014-08-05 Becton, Dickinson And Company Density phase separation device
US10343157B2 (en) 2009-05-15 2019-07-09 Becton, Dickinson And Company Density phase separation device
US9079123B2 (en) 2009-05-15 2015-07-14 Becton, Dickinson And Company Density phase separation device
US10376879B2 (en) 2009-05-15 2019-08-13 Becton, Dickinson And Company Density phase separation device
US10413898B2 (en) 2009-05-15 2019-09-17 Becton, Dickinson And Company Density phase separation device
US10456782B2 (en) 2009-05-15 2019-10-29 Becton, Dickinson And Company Density phase separation device
WO2015070273A1 (fr) * 2013-11-14 2015-05-21 Greiner Bio-One Gmbh Dispositif de réception, procédé de production de celui-ci et procédé de séparation d'un mélange
CN105848580A (zh) * 2013-11-14 2016-08-10 格莱恩比奥-奥内有限公司 容纳装置、用于提供所述容纳装置的方法以及用于分离混合物的方法
US9694359B2 (en) 2014-11-13 2017-07-04 Becton, Dickinson And Company Mechanical separator for a biological fluid

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