Leukocyte concentration and separation device, blood collection container, and leukocyte separation method

Abstract

Provided is a blood collection container with which it is possible to obtain a sample in which the recovery rate of leukocytes is high and the intrusion amount of erythrocytes is low.　The blood collection container according to the present invention is a container in which a prescribed amount of blood is collected, the blood collection container comprising a blood collection container body and a blood cell separation member accommodated within the blood collection container body. The specific gravity at 25°C of the blood cell separation member is 1.075-1.093 (inclusive). The blood collection container either: comprises an erythrocyte agglomeration agent accommodated within the blood collection container body (configuration A2); or comprises an osmotic pressure adjustment agent accommodated within the blood collection container body, and is configured such that, when a water-soluble component accommodated within the blood collection container body is dissolved in a prescribed amount of blood collected in the blood collection container and an equivalent amount of physiological saline to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300-500 mOsm / L (inclusive) (configuration B2).

Description

[Technical Field]

[0001] The present invention relates to a leukocyte concentration and separation device for concentrating and separating leukocytes present in a blood-derived sample, a blood collection container capable of concentrating and separating leukocytes, and a method for separating leukocytes using the leukocyte separation device or the blood collection container. [Background technology]

[0002] In clinical testing, specific cells are concentrated and separated from blood, and tests using the separated cells are performed. For example, red blood cells are removed from the blood and white blood cells are separated to observe the morphology of white blood cells, analyze the surface antigens of white blood cells, or examine the chromosomes and genes inside white blood cells.

[0003] Known methods for removing red blood cells from blood and recovering white blood cells include hemolysis and centrifugation using Ficoll. However, these methods require complicated procedures, and the recovery rate of white blood cells varies greatly depending on the skill of the operator.

[0004] Devices that can easily separate white blood cells without depending on the skill of the operator are described in Patent Documents 1 and 2 listed below.

[0005] Patent Document 1 listed below describes a blood collection container containing a gel composition having a specific specific gravity. When blood is collected in this blood collection container and centrifuged, red blood cells settle to the bottom of the gel composition due to the difference in specific gravity, and white blood cells are concentrated and separated to the top of the gel composition.

[0006] Patent Document 2 listed below describes a blood collection container containing a gel composition having a specific specific gravity and a water-soluble density gradient material having a specific gravity greater than that of the gel composition. When blood is collected in this blood collection container and centrifuged, red blood cells settle below the gel composition due to the difference in specific gravity, and white blood cells are concentrated and separated in a layer formed between the gel composition and the plasma layer. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] WO2019 / 131613A1 [Patent Document 2] Japanese Patent Application Publication No. 61-084557 Summary of the Invention [Problem to be solved by the invention]

[0008] In the method described in Patent Document 1, the amount of red blood cells contaminating the white blood cell layer that accumulates on top of the gel composition after centrifugation may be large. If a white blood cell test is performed using a sample that contains a large amount of red blood cells, the test accuracy may decrease, and normal test results may not be obtained.

[0009] Furthermore, although the method described in Patent Document 2 can reduce the amount of red blood cells contaminating the white blood cell layer to some extent, it is difficult to increase the white blood cell recovery rate. Even when a sample with a low white blood cell recovery rate is used, the test accuracy may decrease, and normal test results may not be obtained.

[0010] An object of the present invention is to provide a leukocyte concentration and separation device and a blood collection container that can obtain a sample with a high recovery rate of leukocytes and a low amount of contaminating red blood cells. Another object of the present invention is to provide a method for separating leukocytes using the leukocyte concentration and separation device or blood collection container. [Means for solving the problem]

[0011] According to a broad aspect of the present invention, there is provided a leukocyte concentration and separation device to which a predetermined amount of blood-derived sample is added, for concentrating and separating leukocytes present in the blood-derived sample, the leukocyte concentration and separation device comprising a container body and a blood cell separation material contained within the container body, the blood cell separation material having a specific gravity of 1.075 or more and 1.093 or less at 25°C, and comprising the following configuration A1 or configuration B1:

[0012] Configuration A1: A red blood cell agglutinating agent is contained in the container body.

[0013] Configuration B1: The device is provided with an osmotic pressure adjusting agent contained within the container body, and when the water-soluble components contained within the container body are dissolved in a volume of physiological saline equal to the predetermined volume of the blood-derived sample to be added to the leukocyte concentration and separation device to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less.

[0014] According to a broad aspect of the present invention, there is provided a blood collection container for collecting a predetermined amount of blood, comprising a blood collection container body and a blood cell separation material contained within the blood collection container body, wherein the specific gravity of the blood cell separation material at 25°C is 1.075 or more and 1.093 or less, and the blood collection container has the following configuration A2 or configuration B2:

[0015] Configuration A2: A red blood cell agglutinating agent is contained within the blood collection container body.

[0016] Configuration B2: The blood collection container body is provided with an osmotic pressure adjusting agent, and when a solution for measuring osmotic pressure is obtained by dissolving the water-soluble component contained in the blood collection container body with a volume of physiological saline equal to the predetermined volume of blood to be collected in the blood collection container, the osmotic pressure of the solution for measuring osmotic pressure is 300 mOsm / L or more and 500 mOsm / L or less.

[0017] In a specific aspect of the blood collection container according to the present invention, the blood cell separation material is a blood cell separation composition having thixotropy.

[0018] In a specific aspect of the blood collection container according to the present invention, the blood collection container has the structure A2.

[0019] In a specific aspect of the blood collection container according to the present invention, the red blood cell agglutinating agent is dextran, hydroxyethyl starch, or polyethylene glycol.

[0020] In a specific aspect of the blood collection container according to the present invention, the dextran has a weight-average molecular weight of 50,000 or more.

[0021] In a specific aspect of the blood collection container according to the present invention, the blood collection container has the structure B2.

[0022] In a specific aspect of the blood collection container according to the present invention, the osmotic pressure of the solution for measuring osmotic pressure is 320 mOsm / L or more and 360 mOsm / L or less.

[0023] In a specific aspect of the blood collection container of the present invention, the specific gravity of the blood cell separation material at 25°C is 1.082 or more and 1.090 or less, and the osmotic pressure of the osmotic pressure measurement solution is 320 mOsm / L or more and 360 mOsm / L or less.

[0024] In a specific aspect of the blood collection container according to the present invention, the osmotic pressure adjusting agent is sodium chloride or glucose.

[0025] In a specific aspect of the blood collection container according to the present invention, the blood collection container comprises the configuration A2 and the configuration B2.

[0026] In a specific aspect of the blood collection container according to the present invention, the blood collection container includes an anticoagulant contained within the blood collection container body.

[0027] In a specific aspect of the blood collection container according to the present invention, the blood collection container includes an antioxidant contained within the blood collection container body.

[0028] In a specific aspect of the blood collection container according to the present invention, the antioxidant is ascorbic acid or an inorganic salt of ascorbic acid.

[0029] According to a broad aspect of the present invention, there is provided a method for separating leukocytes using the above-mentioned leukocyte enrichment and separation device, comprising the steps of adding a blood-derived sample to the leukocyte enrichment and separation device, and centrifuging the leukocyte enrichment and separation device to which the blood-derived sample has been added.

[0030] According to a broad aspect of the present invention, there is provided a method for separating white blood cells using the above-mentioned blood collection container, comprising the steps of collecting blood in the blood collection container and centrifuging the blood collection container in which the blood has been collected.

[0031] According to a broad aspect of the present invention, there is provided a leukocyte concentration and separation system for collecting a predetermined amount of blood-derived sample and concentrating and separating leukocytes present in the blood-derived sample, the leukocyte concentration and separation system comprising a blood cell separation material having a specific gravity at 25°C of 1.075 or more and 1.093 or less, and comprising the following configuration A3 or configuration B3:

[0032] Configuration A3: Erythrocyte agglutinating agent is included.

[0033] Configuration B3: An osmotic pressure adjuster is provided, and when a water-soluble component to be mixed with the blood-derived sample is dissolved in an amount of physiological saline equal to a predetermined amount of the blood-derived sample to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less. [Effects of the Invention]

[0034] The leukocyte concentration and separation device according to the present invention is a leukocyte concentration and separation device to which a predetermined amount of blood-derived sample is added, for concentrating and separating leukocytes present in the blood-derived sample, and comprises a container body and a blood cell separation material contained in the container body, the specific gravity of the blood cell separation material at 25°C being 1.075 or more and 1.093 or less, and is provided with the following configuration A1 or configuration B1. Configuration A1: A red blood cell agglutinating agent is contained in the container body. Configuration B1: An osmotic pressure adjuster is contained in the container body, and when a water-soluble component contained in the container body is dissolved with an amount of physiological saline equal to the predetermined amount of blood-derived sample to be added to the leukocyte concentration and separation device to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less. Because the leukocyte concentration and separation device according to the present invention has the above configuration, it is possible to obtain a sample with a high white blood cell recovery rate and a low amount of red blood cells mixed in.

[0035] The blood collection container according to the present invention is a blood collection container for collecting a predetermined amount of blood, and comprises a blood collection container body and a blood cell separation material contained within the blood collection container body, the specific gravity of the blood cell separation material at 25°C being 1.075 or more and 1.093 or less, and comprises the following configuration A2 or B2. Configuration A2: A red blood cell agglutinating agent is contained within the blood collection container body. Configuration B2: An osmotic pressure adjuster is contained within the blood collection container body, and when a water-soluble component contained within the blood collection container body is dissolved with an amount of physiological saline equal to the predetermined amount of blood to be collected in the blood collection container to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less. Because the blood collection container according to the present invention is provided with the above configurations, it is possible to obtain a sample with a high white blood cell recovery rate and a low amount of red blood cells mixed in. [Brief explanation of the drawings]

[0036] [Figure 1] FIG. 1 is a front cross-sectional view of a blood collection container according to a first embodiment of the present invention. [Figure 2]FIG. 2 is a front cross-sectional view of a blood collection container according to a second embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0037] The present invention will be described in detail below.

[0038] The leukocyte concentration and separation device according to the present invention is a leukocyte concentration and separation device to which a predetermined amount of blood-derived sample is added, for concentrating and separating leukocytes present in the blood-derived sample, and comprises a container body and a blood cell separation material contained within the container body, the specific gravity of the blood cell separation material at 25°C being 1.075 or more and 1.093 or less, and comprising the following configuration A1 or configuration B1:

[0039] Configuration A1: A red blood cell agglutinating agent is contained in the container body.

[0040] Configuration B1: The device is provided with an osmotic pressure adjusting agent contained within the container body, and when the water-soluble components contained within the container body are dissolved in an amount of physiological saline equal to the predetermined amount of blood-derived sample to be added to the leukocyte concentration and separation device to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less.

[0041] The blood collection container of the present invention is a blood collection container for collecting a predetermined amount of blood, and comprises a blood collection container body and a blood cell separation material contained within the blood collection container body, wherein the specific gravity of the blood cell separation material at 25°C is 1.075 or more and 1.093 or less, and comprises the following configuration A2 or configuration B2.

[0042] Configuration A2: A red blood cell agglutinating agent is contained within the blood collection container body.

[0043] Configuration B2: The blood collection container body is provided with an osmotic pressure adjusting agent, and when the water-soluble component contained in the blood collection container body is dissolved in a volume of physiological saline equal to the predetermined volume of blood to be collected in the blood collection container to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less.

[0044] The leukocyte concentration and separation device and blood collection container according to the present invention are provided with the above-described configuration, making it possible to obtain a sample with a high white blood cell recovery rate and a low amount of contaminated red blood cells.The leukocyte concentration and separation device and blood collection container according to the present invention are provided with the above-described configuration, making it possible to obtain a sample with a high white blood cell content and a low red blood cell content.

[0045] The leukocyte enrichment and separation device according to the present invention may include the above-mentioned configuration A1, the above-mentioned configuration B1, or the above-mentioned configuration A1 and the above-mentioned configuration B1. From the viewpoint of more effectively exerting the effects of the present invention, it is preferable that the leukocyte enrichment and separation device according to the present invention includes the above-mentioned configuration A1 and the above-mentioned configuration B1.

[0046] The blood collection container according to the present invention may include the above-mentioned configuration A2, the above-mentioned configuration B2, or the above-mentioned configuration A2 and the above-mentioned configuration B2. From the viewpoint of more effectively exerting the effects of the present invention, it is preferable that the blood collection container according to the present invention includes the above-mentioned configuration A2 and the above-mentioned configuration B2.

[0047] When the blood collection container according to the present invention has the above-described configuration A2, when blood is collected in the blood collection container, red blood cells are agglutinated by the action of the red blood cell agglutinant. By centrifuging the blood collection container, the agglutinated red blood cells move smoothly downward relative to the blood cell separation material having a specific specific gravity. Furthermore, white blood cells move smoothly upward relative to the blood cell separation material having a specific specific gravity. As a result, a sample can be obtained with a high white blood cell recovery rate and a low amount of red blood cells mixed in.

[0048] When the blood collection container according to the present invention is equipped with the above-described configuration B2, the osmotic pressure of the blood increases when blood is collected in the blood collection container. Therefore, the water content in white blood cells and the water content in red blood cells move out of the blood cells, increasing the specific gravity of the white blood cells and red blood cells. As a result, the difference between the specific gravity of the white blood cells and the specific gravity of the red blood cells can be increased. By centrifuging the blood collection container, the red blood cells, whose specific gravity has increased, move smoothly downward relative to the blood cell separation material having a specific specific gravity. Furthermore, the white blood cells move smoothly upward relative to the blood cell separation material having a specific specific gravity. As a result, a sample with a high white blood cell recovery rate and a low amount of red blood cells can be obtained.

[0049] The leukocyte concentration and separation device according to the present invention also has a similar mechanism to that described above, making it possible to obtain a sample with a high recovery rate of leukocytes and a low amount of contaminated erythrocytes.

[0050] When the leukocyte concentration and separation device according to the present invention has both the above-mentioned configuration A1 and the above-mentioned configuration B1, and when the blood collection container according to the present invention has both the above-mentioned configuration A2 and the above-mentioned configuration B2, both the effect due to the aggregation of red blood cells and the effect due to the specific gravity are exerted, and the effects of the present invention are exerted even more effectively.

[0051] In the leukocyte concentration and separation device having the above-mentioned configuration B1, when the water-soluble components contained in the container body are dissolved with an amount of physiological saline equal to a predetermined amount of the blood-derived sample to be added to the leukocyte concentration and separation device to obtain an osmotic pressure measurement solution, the osmotic pressure of the obtained osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less.

[0052] In a blood collection container having the above-mentioned configuration B2, when a solution for measuring osmotic pressure is obtained by dissolving the water-soluble component contained in the blood collection container body with an amount of physiological saline equal to the predetermined amount of blood to be collected in the blood collection container, the osmotic pressure of the obtained solution for measuring osmotic pressure is 300 mOsm / L or more and 500 mOsm / L or less.

[0053] Specifically, the solution for measuring osmotic pressure is prepared as follows.

[0054] A predetermined amount of saline solution equivalent to the predetermined amount of blood-derived sample to be added to the leukocyte concentration / separation device or the predetermined amount of blood to be collected in the blood collection container is added to the leukocyte concentration / separation device or the blood collection container. For example, for a blood collection container that collects 5 mL of blood, 5 mL of saline solution is added to the blood collection container. After addition, the solution is mixed by inversion to dissolve the water-soluble components with the saline solution, thereby obtaining a solution for measuring osmotic pressure. The water-soluble components include, for example, an osmotic pressure adjuster, a red blood cell agglutinating agent, and a liquid containing these.

[0055] The osmotic pressure of the osmotic pressure measurement solution is measured by the freezing point depression method using an osmometer (for example, "OM-6060" manufactured by Arkray).

[0056] The osmotic pressure of the osmotic pressure measurement solution is preferably 300 mOsm / L or more, more preferably 310 mOsm / L or more, even more preferably 320 mOsm / L or more, and preferably 500 mOsm / L or less, more preferably 400 mOsm / L or less, and even more preferably 360 mOsm / L or less. When the osmotic pressure is equal to or greater than the lower limit, red blood cells can be more effectively prevented from being mixed into the sample. When the specific gravity of the blood cell separation material at 25°C is 1.082 or more and 1.090 or less, and the osmotic pressure of the osmotic pressure measurement solution is 320 mOsm / L or more and 360 mOsm / L or less, the effects of the present invention are more effectively exhibited.

[0057] (blood cell separation material) The leukocyte concentration and separation device comprises a blood cell separation material housed in the container body. The blood collection container comprises a blood cell separation material housed in the blood collection container body. The specific gravity of the blood cell separation material at 25°C is 1.075 or more and 1.093 or less. Examples of the blood cell separation material include a blood cell separation composition and a blood cell separation tool. Because the blood cell separation material is easy to prepare, the blood cell separation material is preferably the blood cell separation composition.

[0058] The location where the blood cell separation material is contained is not particularly limited as long as it is within the container body or the blood collection container body. In the leukocyte concentration and separation device, the blood cell separation material may be disposed at the bottom of the container body or on the inner wall surface. In the blood collection container, the blood cell separation material may be disposed at the bottom of the blood collection container body or on the inner wall surface.

[0059] <Composition for blood cell separation> The blood cell separation composition is a composition that migrates between white blood cells and red blood cells during centrifugation to form a partition. The blood cell separation composition is used for the purpose of preventing red blood cells from contaminating the white blood cell layer after centrifugation. The blood cell separation composition is preferably thixotropic, and more preferably a thixotropic gel-like composition. In the white blood cell concentration and separation device, the blood cell separation composition may be contained in the bottom of the container body or may be disposed on the inner wall surface. From the viewpoint of more effectively achieving the effects of the present invention, the blood cell separation composition is preferably contained in the bottom of the container body. In the blood collection container, the blood cell separation composition may be contained in the bottom of the blood collection container body or may be disposed on the inner wall surface. From the viewpoint of more effectively achieving the effects of the present invention, the blood cell separation composition is preferably contained in the bottom of the blood collection container body.

[0060] The composition for separating blood cells preferably contains an organic component that is fluid at 25° C. and an inorganic fine powder. The organic component that is fluid at 25° C. and the inorganic fine powder may each be used alone or in combination of two or more types.

[0061] Organic components that are flowable at 25°C: The above phrase "having fluidity at 25°C" means that the viscosity at 25°C is 500 Pa·s or less.

[0062] The viscosity of the organic component at 25°C is preferably 30 Pa s or more, more preferably 50 Pa s or more, and preferably 200 Pa s or less, more preferably 100 Pa s or less. When the viscosity is equal to or greater than the above lower limit and equal to or less than the above upper limit, the fluidity of the composition for separating blood cells is increased, and the strength of the partition wall can be increased.

[0063] The viscosity of the organic component at 25°C was measured using an E-type viscometer (for example, "TVE-35" manufactured by Toki Sangyo Co., Ltd.) at 25°C and a shear rate of 1.0 sec. -1 It is measured under the following conditions.

[0064] Examples of the organic component include a resin and a mixture of a resin and an organic compound such as a plasticizer. When the organic component is a mixture of the resin and the organic compound, the mixture (the organic component) needs to have fluidity, and the resin or the organic compound does not need to have fluidity. When the organic component is a mixture of the resin and the organic compound, the resin may be, for example, a resin that is solid at 25°C. Only one type of the resin and the organic compound may be used, or two or more types may be used in combination.

[0065] Examples of the resin include petroleum resins, cyclopentadiene resins, polyester resins, polyurethane resins, (meth)acrylic resins, silicone resins, α-olefin-fumaric acid ester copolymers, copolymers of sebacic acid, 2,2-dimethyl-1,3-propanediol, and 1,2-propanediol, polyether polyurethane resins, and polyether polyester resins, etc. Only one of the resins may be used, or two or more of them may be used in combination.

[0066] Commercially available petroleum resins include "Regalite S5090" manufactured by Eastman Chemical Company.

[0067] Examples of the cyclopentadiene resin include a polymer of a cyclopentadiene monomer, a copolymer of a cyclopentadiene monomer and an aromatic monomer, and a dicyclopentadiene resin. The cyclopentadiene resin may be hydrogenated. The polymer of a cyclopentadiene monomer and the copolymer of a cyclopentadiene monomer and an aromatic monomer may be an oligomer.

[0068] Examples of the cyclopentadiene-based monomer include cyclopentadiene, dicyclopentadiene, and alkyl-substituted derivatives of cyclopentadiene.

[0069] Examples of the aromatic monomer include styrene, methylstyrene, indene, and methylindene.

[0070] Commercially available dicyclopentadiene resins include "Scoretz SU500" and "Scoretz SU90" manufactured by Colon Co., Ltd.

[0071] Examples of the polyester resin include polyalkylene terephthalate resins and polyalkylene naphthalate resins, etc. Examples of the polyalkylene terephthalate resin include polyethylene terephthalate, polybutylene terephthalate, and poly-1,4-cyclohexanedimethylene terephthalate.

[0072] Examples of the polyurethane resin include a reaction product of a polyol compound and an isocyanate compound.

[0073] Examples of the (meth)acrylic resin include a resin obtained by polymerizing at least one type of (meth)acrylic acid ester monomer, and a resin obtained by polymerizing at least one type of (meth)acrylic acid ester monomer and at least one type of monomer other than the (meth)acrylic acid ester monomer.

[0074] Examples of the (meth)acrylic acid ester monomer include (meth)acrylic acid alkyl esters having an alkyl group having from 1 to 20 carbon atoms, (meth)acrylic acid polyalkylene glycol esters, (meth)acrylic acid alkoxyalkyl esters, (meth)acrylic acid hydroxyalkyl esters, (meth)acrylic acid glycidyl esters, (meth)acrylic acid dialkylaminoalkyl esters, (meth)acrylic acid benzyl esters, (meth)acrylic acid phenoxyalkyl esters, (meth)acrylic acid cyclohexyl esters, (meth)acrylic acid isobornyl esters, and (meth)acrylic acid alkoxysilyl alkyl esters. The above (meth)acrylic acid ester monomers may be used alone or in combination of two or more.

[0075] The organic compound may be a benzene polycarboxylic acid alkyl ester derivative. The organic compound is preferably a benzene polycarboxylic acid alkyl ester derivative. Therefore, the organic component is preferably a mixture of the resin and the benzene polycarboxylic acid alkyl ester derivative.

[0076] Examples of the benzene polycarboxylic acid alkyl ester derivatives include phthalates, trimellitates, pyromellitates, etc. The benzene polycarboxylic acid alkyl ester derivatives may be used alone or in combination of two or more.

[0077] Examples of the trimellitic acid ester include tri-n-octyl trimellitate, triisooctyl trimellitate, and triisodecyl trimellitate.

[0078] Examples of the pyromellitic acid ester include tetraisooctyl pyromellitic acid.

[0079] Commercially available trimellitic acid esters include "Monocizer W700" and "Monocizer W-750" manufactured by DIC Corporation, and "Sansocizer TO™" and "Sansocizer TI™" manufactured by New Japan Chemical Co., Ltd.

[0080] Commercially available pyromellitic acid esters include "Monocizer W-7010" manufactured by DIC Corporation.

[0081] The benzenepolycarboxylic acid alkyl ester derivative is preferably a phthalate ester, a trimellitate ester, or a pyromellitate ester, and more preferably a trimellitate ester.

[0082] The content of the organic component in 100% by weight of the composition for separating blood cells is preferably 80% by weight or more, more preferably 85% by weight or more, even more preferably 90% by weight or more, and preferably 95% by weight or less.

[0083] Inorganic fine powder: Examples of the inorganic fine powder include fine silica powder, titanium oxide powder, zinc oxide powder, calcium carbonate powder, alumina powder, fine glass powder, talc powder, kaolin powder, bentonite powder, titania powder, and zirconium powder.

[0084] The inorganic fine powder preferably contains fine silica and an inorganic fine powder different from the fine silica. The inorganic fine powder different from the fine silica is preferably an inorganic fine powder having a higher specific gravity than the fine silica, and more preferably an inorganic fine powder having a specific gravity of 3 or more, such as zinc oxide powder, titanium oxide powder, or alumina powder.

[0085] In order to more effectively exert the effects of the present invention, the inorganic fine powder preferably contains finely powdered silica.

[0086] The finely powdered silica includes natural silica and synthetic silica. Synthetic silica includes hydrophilic silica and hydrophobic silica. Hydrophilic silica imparts thixotropy to the composition for blood cell separation and adjusts the specific gravity by hydrogen bonding between hydroxyl groups on the particle surface. On the other hand, hydrophobic silica has a smaller effect of imparting thixotropy than hydrophilic silica.

[0087] From the viewpoint of maintaining both the specific gravity and thixotropy of the composition for separating blood cells within a suitable range, the finely powdered silica preferably contains hydrophilic silica, and more preferably contains hydrophilic silica and hydrophobic silica. The finely powdered silica preferably contains at least hydrophilic silica.

[0088] The content of hydrophilic silica in 100% by weight of the composition for blood cell separation is preferably 0.01% by weight or more, more preferably 0.1% by weight or more, even more preferably 0.3% by weight or more, and preferably 2.50% by weight or less, more preferably 2.00% by weight or less. When the content of hydrophilic silica is equal to or more than the lower limit and equal to or less than the upper limit, both the specific gravity and thixotropy of the composition for blood cell separation can be maintained within more suitable ranges.

[0089] The content of finely powdered silica in 100% by weight of the composition for blood cell separation is preferably 0.1% by weight or more, more preferably 1% by weight or more, even more preferably 3% by weight or more, and preferably 10% by weight or less, more preferably 5% by weight or less. When the content of finely powdered silica is equal to or more than the above lower limit and equal to or less than the above upper limit, both the specific gravity and thixotropy of the composition for blood cell separation can be maintained within more suitable ranges.

[0090] The average particle size of the finely powdered silica and the inorganic fine powder different from the finely powdered silica is not particularly limited. The average particle size of the finely powdered silica may be 1 nm or more, 10 nm or more, 500 nm or less, or 100 nm or less. The average particle size of the inorganic fine powder different from the finely powdered silica is not particularly limited. The average particle size of the inorganic fine powder may be 10 nm or more, 100 nm or more, and 10 μm or less, or 1 μm or less.

[0091] The average particle diameter of the finely powdered silica and the inorganic fine powder different from the finely powdered silica is the average diameter measured on a volume basis, and is the value of the 50% median diameter (D50). The volume average particle diameter (D50) can be measured by laser diffraction / scattering method, image analysis method, Coulter method, centrifugal sedimentation method, etc. The volume average particle diameter (D50) is preferably determined by measurement using laser diffraction / scattering method or image analysis method.

[0092] The content of the inorganic fine powder in 100% by weight of the composition for blood cell separation is preferably 0.1% by weight or more, more preferably 1% by weight or more, even more preferably 3% by weight or more, and preferably 10% by weight or less, more preferably 5% by weight or less. When the content of the inorganic fine powder is equal to or more than the above lower limit and equal to or less than the above upper limit, both the specific gravity and thixotropy of the composition for blood cell separation can be maintained within more suitable ranges.

[0093] Other Ingredients: The composition for separating blood cells may contain other components in addition to the components described above, as long as the effects of the present invention are not impaired. The composition for separating blood cells may contain, for example, an organic gelling agent, a thermoplastic elastomer, a polyalkylene glycol, a silicone oil, a cosolvent, an antioxidant, a colorant, water, etc. Each of the other components may be used alone, or two or more may be used in combination.

[0094] Other details: The specific gravity of the composition for separating blood cells at 25° C. is 1.075 or more and 1.093 or less. The specific gravity of the composition for separating blood cells at 25° C. is preferably 1.077 or more, more preferably 1.082 or more, preferably 1.090 or less, and more preferably 1.088 or less. When the specific gravity is equal to or more than the lower limit and equal to or less than the upper limit, the recovery rate of white blood cells is further increased, and a sample with even less red blood cell contamination can be successfully obtained.

[0095] The specific gravity of the above-mentioned blood cell separation composition at 25°C is measured by dropping one drop of the blood cell separation composition into saline solution at 25°C, the specific gravity of which has been adjusted in steps at intervals of 0.002, and observing whether the composition floats or sinks in the saline solution.

[0096] The viscosity of the composition for separating blood cells at 25° C. is preferably 100 Pa s or more, more preferably 150 Pa s or more, and preferably 500 Pa s or less, more preferably 400 Pa s or less. When the viscosity is equal to or more than the above lower limit and equal to or less than the above upper limit, the effects of the present invention can be more effectively exhibited.

[0097] The viscosity of the composition for blood cell separation at 25°C was measured using an E-type viscometer (for example, "TVE-35" manufactured by Toki Sangyo Co., Ltd.) at 25°C and a shear rate of 1.0 sec -1 It is measured under the following conditions.

[0098] <Blood cell separation tool> The blood cell separation device is a device that moves between white blood cells and red blood cells during centrifugation to form a partition wall, and is used for the purpose of preventing red blood cells from mixing into the white blood cell layer after centrifugation.

[0099] The blood cell separation device may be, for example, a device known as a serum or plasma separation device, such as the mechanical separator (blood cell separation device) described in WO2010 / 132783A1.

[0100] Examples of materials for the blood cell separation tool include elastomers.

[0101] (red blood cell agglutinating agent) The leukocyte concentration / separation device having the above-mentioned configuration A1 includes a red blood cell agglutinating agent contained in the container body. The blood collection container having the above-mentioned configuration A2 includes a red blood cell agglutinating agent contained in the blood collection container body. A conventionally known red blood cell agglutinating agent can be used as the red blood cell agglutinating agent. Only one type of the red blood cell agglutinating agent may be used, or two or more types may be used in combination.

[0102] Examples of the red blood cell agglutinating agents include polysaccharides such as dextran, hydroxyethyl starch, methylcellulose, and polysucrose; cationic polymers such as polyamino acids, polyamine sulfones, polybrene, polyalkylene oxides, polyalkyleneimines, polyacrylamides, and cationically modified polyvinyl alcohols; polyethylene glycol (PEG); low molecular weight compounds such as vancomycin and kanamycin; and cell-binding glycoproteins such as plant lectins.

[0103] The red blood cell agglutinating agent is preferably a polysaccharide or polyethylene glycol, more preferably a polysaccharide. The red blood cell agglutinating agent is preferably dextran, hydroxyethyl starch, or polyethylene glycol, more preferably dextran. In these cases, the effects of the present invention are more effectively exhibited. In addition, the adhesion between the blood cell separation composition and the container body or blood collection container body is improved, and red blood cells can be effectively prevented from remaining in the gap between the blood cell separation composition and the container body or blood collection container body after centrifugation.

[0104] The weight-average molecular weight of the dextran is preferably 50,000 or more, more preferably 70,000 or more, even more preferably 100,000 or more, preferably 1,000,000 or less, more preferably 600,000 or less, and even more preferably 300,000 or less. When the weight-average molecular weight is equal to or greater than the above-mentioned lower limit and equal to or less than the above-mentioned upper limit, the effects of the present invention are even more effectively exhibited. In addition, the adhesion between the blood cell separation composition and the container body or blood collection container body is further improved, and red blood cells can be even more effectively prevented from remaining in the gap between the blood cell separation composition and the container body or blood collection container body after centrifugation.

[0105] The weight-average molecular weight of the hydroxyethyl starch is preferably 70,000 or more, more preferably 100,000 or more, even more preferably 200,000 or more, and preferably 1,000,000 or less, more preferably 700,000 or less, and even more preferably 500,000 or less. When the weight-average molecular weight is equal to or greater than the above-mentioned lower limit and equal to or less than the above-mentioned upper limit, the effects of the present invention are even more effectively exhibited. In addition, the adhesion between the blood cell separation composition and the container body or blood collection container body is further improved, and red blood cells can be even more effectively prevented from remaining in the gap between the blood cell separation composition and the container body or blood collection container body after centrifugation.

[0106] The weight-average molecular weight of the polyethylene glycol is preferably 1,000 or more, more preferably 3,000 or more, even more preferably 5,000 or more, and preferably 100,000 or less, more preferably 50,000 or less, and even more preferably 30,000 or less. When the weight-average molecular weight is equal to or greater than the above-mentioned lower limit and equal to or less than the above-mentioned upper limit, the effects of the present invention are even more effectively exhibited. In addition, the adhesion between the blood cell separation composition and the container body or blood collection container body is further improved, and red blood cells can be even more effectively prevented from remaining in the gap between the blood cell separation composition and the container body or blood collection container body after centrifugation.

[0107] The weight average molecular weight is measured by gel permeation chromatography (GPC) and is expressed as pullulan.

[0108] In the leukocyte concentration / separation device, the erythrocyte agglutinating agent may be contained in the container body in a powder state or in a dissolved state in liquid. When the erythrocyte agglutinating agent is contained in the container body in a dissolved state in liquid, it is preferable that the osmotic pressure of the liquid is adjusted to be isotonic with blood in order to prevent hemolysis when blood flows in. The erythrocyte agglutinating agent may be present in the container body in both a powder state and a dissolved state in liquid.

[0109] In the blood collection container, the red blood cell agglutinating agent may be contained in the blood collection container body in a powdered state or in a dissolved state in liquid. When the red blood cell agglutinating agent is contained in the blood collection container body in a dissolved state in liquid, it is preferable that the osmotic pressure of the liquid is adjusted to be isotonic with blood in order to prevent hemolysis when blood is introduced. The red blood cell agglutinating agent may be present in the blood collection container body in both a powdered state and a dissolved state in liquid.

[0110] The liquid may be water, alcohol, or the like.

[0111] The red blood cell agglutinating agent is preferably disposed on the inner wall surface of the blood collection container body or on the surface of the blood cell separation material. The red blood cell agglutinating agent may be disposed on the inner wall surface of the blood collection container body, on the surface of the blood cell separation material, or on both the inner wall surface of the blood collection container body and the surface of the blood cell separation material. Similarly, in the case of the leukocyte concentration separation device, the red blood cell agglutinating agent is preferably disposed on the inner wall surface of the container body or on the surface of the blood cell separation material.

[0112] When the red blood cell agglutinating agent is contained in a powdered state, the powdered red blood cell agglutinating agent is preferably attached to the inner wall surface of the blood collection container body or disposed on the surface of the blood cell separation material.Similarly, in the case of the leukocyte concentration separation device, the powdered red blood cell agglutinating agent is preferably attached to the inner wall surface of the container body or disposed on the surface of the blood cell separation material.

[0113] In the leukocyte concentration / separation device, the content of the erythrocyte agglutinating agent is preferably 5 mg or more, more preferably 10 mg or more, preferably 50 mg or less, and more preferably 30 mg or less per mL of blood-derived sample added. In the blood collection container, the content of the erythrocyte agglutinating agent is preferably 5 mg or more, more preferably 10 mg or more, preferably 50 mg or less, and more preferably 30 mg or less per mL of blood collected. When the content of the erythrocyte agglutinating agent is above the above-mentioned lower limit and below the above-mentioned upper limit, the effects of the present invention are even more effectively exhibited. In addition, the adhesion between the blood cell separation composition and the blood collection container body is further improved, and the remaining of erythrocytes in the gap between the blood cell separation composition and the blood collection container body after centrifugation can be even more effectively suppressed.

[0114] (osmolality adjuster) The leukocyte concentration / separation device having the above-mentioned configuration B1 includes an osmotic pressure adjuster housed in the container body. The blood collection container having the above-mentioned configuration B2 includes an osmotic pressure adjuster housed in the blood collection container body. As the osmotic pressure adjuster, a conventionally known osmotic pressure adjuster can be used. Only one type of the osmotic pressure adjuster may be used, or two or more types may be used in combination.

[0115] Examples of the osmotic pressure adjuster include sodium chloride, potassium chloride, glucose, dihydroxyacetone, and sugar alcohols such as D-mannitol and D-sorbitol.

[0116] In order to more effectively exert the effects of the present invention, the osmotic pressure adjusting agent is preferably sodium chloride or glucose.

[0117] In the leukocyte concentration / separation device, the osmotic pressure adjuster may be contained in the container body in the form of a powder or in the form of a solution in a liquid. Note that the osmotic pressure adjuster may be present in the container body in both the form of a powder and the form of a solution in a liquid.

[0118] In the blood collection container, the osmotic pressure adjuster may be contained in the blood collection container body in the form of a powder or in the form of a solution in a liquid, or the osmotic pressure adjuster may be present in the blood collection container body in both the form of a powder and the form of a solution in a liquid.

[0119] The liquid may be water, alcohol, or the like.

[0120] The osmotic pressure adjuster is preferably disposed on the inner wall surface of the blood collection container body or on the surface of the blood cell separation material. The osmotic pressure adjuster may be disposed on the inner wall surface of the blood collection container body, on the surface of the blood cell separation material, or on both the inner wall surface of the blood collection container body and the surface of the blood cell separation material. Similarly, in the case of the leukocyte concentration separation device, the osmotic pressure adjuster is preferably disposed on the inner wall surface of the container body or on the surface of the blood cell separation material.

[0121] When the osmotic pressure adjuster is contained in a powdered state, the powdered osmotic pressure adjuster is preferably attached to the inner wall surface of the blood collection container body or disposed on the surface of the blood cell separation material.Similarly, in the case of the leukocyte concentration separation device, the powdered osmotic pressure adjuster is preferably attached to the inner wall surface of the container body or disposed on the surface of the blood cell separation material.

[0122] The amount of the osmotic pressure adjusting agent contained in the container body or the blood collection container body is not particularly limited as long as the osmotic pressure of the osmotic pressure measuring solution satisfies the above-mentioned range.

[0123] (anticoagulant) The leukocyte concentration / separation device preferably includes an anticoagulant housed in the container body. The blood collection container preferably includes an anticoagulant housed in the blood collection container body. A conventionally known anticoagulant can be used as the anticoagulant. Only one type of anticoagulant may be used, or two or more types may be used in combination.

[0124] Examples of the anticoagulant include heparin, ethylenediaminetetraacetic acid (EDTA), and citric acid.

[0125] In the leukocyte concentration / separation device, the anticoagulant may be contained in the container body in a powder state or in a dissolved state in liquid. When the anticoagulant is contained in the container body in a dissolved state in liquid, it is preferable that the osmotic pressure of the liquid is adjusted to be isotonic with blood in order to prevent hemolysis when blood flows in. The anticoagulant may be present in the container body in both a powder state and a dissolved state in liquid.

[0126] In the blood collection container, the anticoagulant may be contained in the blood collection container body in a powdered state or in a dissolved state in a liquid. If the anticoagulant is contained in the blood collection container body in a dissolved state in a liquid, it is preferable that the osmotic pressure of the liquid be adjusted to be isotonic with blood in order to prevent hemolysis when blood is introduced. The anticoagulant may be present in the blood collection container body in both a powdered state and a dissolved state in a liquid.

[0127] The liquid may be water, alcohol, or the like.

[0128] The anticoagulant is preferably disposed on the inner wall surface of the blood collection container body or on the surface of the blood cell separation material. The anticoagulant may be disposed on the inner wall surface of the blood collection container body, on the surface of the blood cell separation material, or on both the inner wall surface of the blood collection container body and the surface of the blood cell separation material. Similarly, in the case of the leukocyte concentration separation device, the anticoagulant is preferably disposed on the inner wall surface of the container body or on the surface of the blood cell separation material.

[0129] When the anticoagulant is contained in a powdered state, the powdered anticoagulant is preferably attached to the inner wall surface of the blood collection container body or disposed on the surface of the blood cell separation material.Similarly, in the case of the leukocyte concentration and separation device, the powdered anticoagulant is preferably attached to the inner wall surface of the container body or disposed on the surface of the blood cell separation material.

[0130] The amount of the anticoagulant contained in the container body or the blood collection container body is not particularly limited as long as it does not impair the effects of the present invention.

[0131] (antioxidant) The leukocyte concentration / separation device preferably comprises an antioxidant contained within the container body. The blood collection container preferably comprises an antioxidant contained within the blood collection container body. By providing the antioxidant, denaturation of the leukocyte concentration / separation device and the blood collection container during radiation sterilization can be effectively suppressed. Conventionally known antioxidants can be used as the antioxidant. Only one type of antioxidant may be used, or two or more types may be used in combination.

[0132] Examples of the antioxidant include ascorbic acid and inorganic salts of ascorbic acid.

[0133] In the leukocyte enrichment / separation device, the antioxidant may be contained in the container body in a powder state or in a dissolved state in a liquid. When the antioxidant is contained in the container body in a dissolved state in a liquid, it is preferable that the osmotic pressure of the liquid is adjusted to be isotonic with blood in order to prevent hemolysis when blood flows in. The antioxidant may be present in the container body in both a powder state and a dissolved state in a liquid.

[0134] In the blood collection container, the antioxidant may be contained in the blood collection container body in a powder state or in a dissolved state in a liquid. When the antioxidant is contained in the blood collection container body in a dissolved state in a liquid, it is preferable that the osmotic pressure of the liquid is adjusted to be isotonic with blood in order to prevent hemolysis when blood is introduced. The antioxidant may be present in the blood collection container body in both a powder state and a dissolved state in a liquid.

[0135] The liquid may be water, alcohol, or the like.

[0136] The antioxidant is preferably disposed on the inner wall surface of the blood collection container body or on the surface of the blood cell separation material. The antioxidant may be disposed on the inner wall surface of the blood collection container body, on the surface of the blood cell separation material, or on both the inner wall surface of the blood collection container body and the surface of the blood cell separation material. Similarly, in the case of the leukocyte concentration separation device, the antioxidant is preferably disposed on the inner wall surface of the container body or on the surface of the blood cell separation material.

[0137] When the antioxidant is contained in a powdered state, the powdered antioxidant is preferably attached to the inner wall surface of the blood collection container body or disposed on the surface of the blood cell separation material.Similarly, in the case of the leukocyte concentration and separation device, the powdered antioxidant is preferably attached to the inner wall surface of the container body or disposed on the surface of the blood cell separation material.

[0138] The amount of the antioxidant contained in the container body or the blood collection container body is not particularly limited as long as it does not impair the effects of the present invention.

[0139] (Container body and blood collection container body) The shape of the container body and the blood collection container body is not particularly limited, but it is preferable that they are tubular containers with a bottom.

[0140] The material of the container body and the blood collection container body is not particularly limited. Examples of materials for the container body and the blood collection container body include thermoplastic resins such as polyethylene, polypropylene, polystyrene, polyethylene terephthalate, polymethyl methacrylate, and polyacrylonitrile; thermosetting resins such as unsaturated polyester resin, epoxy resin, and epoxy-acrylate resin; modified natural resins such as cellulose acetate, cellulose propionate, ethyl cellulose, and ethyl chitin; and glass such as silicate glass, such as soda-lime glass, phosphosilicate glass, and borosilicate glass, and quartz glass. The container body and the blood collection container body may be made of one material or two or more materials in combination.

[0141] (stopper) The leukocyte concentration / separation device and the blood collection container preferably include a stopper. A conventionally known stopper can be used as the stopper. The stopper is preferably made of a material and has a shape that allows it to be attached to the opening of the container body or the opening of the blood collection container body in an airtight and liquid-tight manner. The stopper is preferably configured to be pierceable by a blood collection needle.

[0142] Examples of the stopper include a stopper having a shape that fits into the opening of the container body or the opening of the blood collection container body, a sheet-like sealing stopper, and the like.

[0143] The stopper may also comprise a stopper body such as a rubber stopper and a cap member made of plastic, etc. In this case, the risk of blood coming into contact with the human body can be reduced when the stopper body is pulled out from the opening of the blood collection container body after blood collection.

[0144] Examples of materials for the stopper (or the stopper main body) include synthetic resin, elastomer, rubber, and metal foil. Examples of the rubber include butyl rubber and halogenated butyl rubber. Examples of the metal foil include aluminum foil. From the viewpoint of improving sealing performance, the stopper is preferably made of butyl rubber. The stopper is preferably a butyl rubber stopper.

[0145] (Other details of blood collection container) The blood collection container is preferably a blood collection tube, and the blood collection container body is preferably a blood collection tube body.

[0146] The blood collection container (a blood collection container having configuration A2 and configuration B2) containing the red blood cell agglutinating agent and the osmotic pressure adjusting agent dissolved in a liquid can be manufactured, for example, as follows.

[0147] The red blood cell agglutinating agent, the osmotic pressure adjusting agent, and other components used as needed are dissolved in a solvent such as water to obtain a mixed solution. The mixed solution obtained is added to the blood collection container body. Furthermore, before or after adding the mixed solution, a blood cell separating composition is placed in the blood collection container body.

[0148] Furthermore, by volatilizing the solvent in the mixed liquid or adding the red blood cell agglutinating agent and the osmotic pressure adjusting agent, etc. in powder form to the surface of the blood cell separation composition, it is possible to obtain the blood collection container in which the red blood cell agglutinating agent and the osmotic pressure adjusting agent are arranged in powder form on the surface of the blood cell separation composition.

[0149] The blood collection container (a blood collection container having configuration A2 and configuration B2) in which the red blood cell agglutinating agent and the osmotic pressure adjusting agent are arranged in powder form on the inner wall surface of the blood collection container body can be manufactured, for example, as follows.

[0150] The red blood cell agglutinating agent, the osmotic pressure adjusting agent, and other components used as needed are dissolved in a solvent such as water to obtain a mixed solution. The mixed solution is applied to the inner wall surface of a blood collection container body and allowed to dry. Furthermore, before or after applying the mixed solution, a blood cell separating composition is placed inside the blood collection container body.

[0151] In the blood collection container, the red blood cell agglutinating agent may be contained in a state dissolved in a liquid, and the osmotic pressure adjuster may be contained in a state of powder. In the blood collection container, for example, the red blood cell agglutinating agent may be contained in a state of powder, and the osmotic pressure adjuster may be contained in a state of dissolved in a liquid.

[0152] Furthermore, by using or not using the red blood cell agglutinating agent and the osmotic pressure adjusting agent, it is possible to manufacture a blood collection container having the structure A2 and a blood collection container having the structure B2.

[0153] FIG. 1 is a front cross-sectional view of a blood collection container according to a first embodiment of the present invention.

[0154] The blood collection container 1 shown in Figure 1 comprises a blood collection container body 2, a blood cell separation composition 3, a mixed solution 4 containing a red blood cell agglutinating agent, an osmotic pressure adjuster, and water, and a stopper 5. The blood collection container body 2 has an opening at one end and a closed bottom at the other end. The blood cell separation composition 3 is contained in the bottom of the blood collection container body 2. The stopper 5 is inserted into the opening of the blood collection container body 2.

[0155] The mixed liquid 4 is placed on the surface of the composition for blood cell separation 3, more specifically, on the upper surface (surface on one end side) of the composition for blood cell separation 3. When the blood collection container is in an upright position, the mixed liquid 4 is placed on the surface of the composition for blood cell separation 3. The red blood cell agglutinating agent and the osmotic pressure adjusting agent are contained in the blood collection container body 2 in a state dissolved in liquid.

[0156] FIG. 2 is a front cross-sectional view of a blood collection container according to a second embodiment of the present invention.

[0157] The blood collection container 1A shown in Figure 2 comprises a blood collection container body 2, a blood cell separation composition 3, a mixed powder 4A of a red blood cell agglutinating agent and an osmotic pressure adjuster, and a stopper 5. The blood collection container body 2 has an opening at one end and a closed bottom at the other end. The blood cell separation composition 3 is contained in the bottom of the blood collection container body 2. The stopper 5 is inserted into the opening of the blood collection container body 2.

[0158] The mixed powder 4A is disposed on the inner wall surface 2a of the blood collection container body. The mixed powder 4A is attached to the inner wall surface 2a of the blood collection container body. That is, the erythrocyte agglutinating agent and the osmotic pressure adjuster are attached to the inner wall surface 2a of the blood collection container body. The erythrocyte agglutinating agent and the osmotic pressure adjuster are contained in the blood collection container body 2 in a powder state. The mixed powder 4A is disposed closer to one end than the composition for blood cell separation 3.

[0159] In the blood collection container according to the present invention, the blood cell separation composition may be disposed on the inner wall surface of the blood collection container body, and the mixed liquid may be disposed at the bottom of the blood collection container body when the blood collection container is in an upright position. Furthermore, in the blood collection container according to the present invention, the blood cell separation composition may be disposed on the inner wall surface of the blood collection container body, and the mixed powder may be disposed on the inner wall surface of the blood collection container body or on the surface of the blood cell separation composition. In the blood collection container according to the present invention, the blood cell separation composition may be disposed on the inner wall surface of the blood collection container body, and the mixed powder may be disposed at the bottom of the blood collection container body. Furthermore, the blood cell separation tool may be used instead of the blood cell separation composition.

[0160] The internal pressure of the leukocyte concentration / separation device and the blood collection container is not particularly limited. The blood collection container can also be used as a vacuum blood collection tube, which is evacuated and then sealed with the sealing member. When used as a vacuum blood collection tube, a fixed amount of blood can be easily collected regardless of the skill level of the blood collector.

[0161] From the viewpoint of preventing bacterial infection, the inside of the leukocyte concentration / separation device and the blood collection container are preferably sterilized in accordance with ISO and JIS standards.

[0162] (Leukocyte separation method) The method for separating white blood cells according to the present invention is a method for separating white blood cells using the blood collection container described above, and includes the steps of collecting blood in the blood collection container and centrifuging the blood collection container with the collected blood.The method for separating white blood cells is a method for separating white blood cells from blood.The method for separating white blood cells is a method for concentrating and separating white blood cells from blood.

[0163] Furthermore, white blood cells can be separated in a similar manner using the white blood cell concentration separation device. The white blood cell separation method includes the steps of adding a blood-derived sample to the white blood cell concentration separation device and centrifuging the white blood cell concentration separation device to which the blood-derived sample has been added.

[0164] In the centrifugal separation step, a partition made of a blood cell separation material can be formed between white blood cells and red blood cells. After centrifugation, white blood cells can be concentrated and separated on the partition formed by the blood cell separation material. After centrifugation, white blood cells are preferably deposited on the partition. The white blood cells concentrated and separated on the partition can be collected and used as a sample.

[0165] The centrifugation conditions in the centrifugal separation step are not particularly limited as long as a partition can be formed using the blood cell separation material to separate white blood cells from red blood cells. Examples of the centrifugation conditions include centrifugation at 400 G or more and 4000 G or less for 10 minutes or more and 120 minutes or less.

[0166] (Leukocyte Separation System) The leukocyte concentration and separation system of the present invention is a leukocyte concentration and separation system in which a predetermined amount of blood-derived sample is collected and the system concentrates and separates leukocytes present in the blood-derived sample, and the system is equipped with a blood cell separation material having a specific gravity at 25°C of 1.075 or more and 1.093 or less, and is equipped with the following configuration A3 or configuration B3.

[0167] Configuration A3: Erythrocyte agglutinating agent is included.

[0168] Configuration B3: An osmotic pressure adjuster is provided, and when a water-soluble component to be mixed with the blood-derived sample is dissolved in an amount of physiological saline equal to a predetermined amount of the blood-derived sample to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 300 mOsm / L or more and 500 mOsm / L or less.

[0169] The leukocyte separation system according to the present invention is provided with the above-described configuration, and therefore can obtain a sample with a high recovery rate of leukocytes and a low amount of contaminated erythrocytes.

[0170] The leukocyte concentration and separation system according to the present invention may include the above-mentioned configuration A3, the above-mentioned configuration B3, or the above-mentioned configuration A3 and the above-mentioned configuration B3. From the viewpoint of more effectively exerting the effects of the present invention, it is preferable that the leukocyte concentration and separation system according to the present invention includes the above-mentioned configuration A3 and the above-mentioned configuration B3.

[0171] The water-soluble component to be mixed with the blood-derived sample in the configuration B3 is, for example, an osmotic pressure adjuster, a red blood cell agglutinating agent, or a liquid containing these.

[0172] In the leukocyte concentration and separation system, the preferred configurations of the erythrocyte agglutinating agent, the osmotic pressure adjusting agent, and the osmotic pressure measuring solution are the same as those described above.

[0173] The leukocyte concentration and separation system preferably includes a storage unit for storing a blood-derived sample. The blood cell separation material, the erythrocyte agglutinating agent, and the osmotic pressure adjusting agent are preferably, but not limited to, stored in the storage unit.

[0174] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples. The following Examples 1 to 8 and 16 are reference examples.

[0175] The following materials were prepared as the composition for blood cell separation.

[0176] (Organic material that is fluid at 25°C) (Meth)acrylic resin: 2-Ethylhexyl acrylate and butyl acrylate were radically polymerized by solution polymerization in the presence of an azo-based polymerization initiator to obtain a (meth)acrylate resin with flowability at 25°C.

[0177] Other resins: Petroleum resin (Eastman Chemical Company's "Rigalite S5090") Dicyclopentadiene resin 1 (Colon "Scolez SU500") Dicyclopentadiene resin 2 (Colon "Scolez SU90")

[0178] Organic compounds: Trimellitic acid ester (benzenepolycarboxylic acid alkyl ester derivative, DIC "Monocizer W700")

[0179] (Inorganic fine powder) Hydrophilic silica (fine powder silica, Nippon Aerosil "200CF") Hydrophobic silica (fine powder silica, Nippon Aerosil "R974") Titanium oxide powder (Ishihara Sangyo Kaisha "A-100")

[0180] (Other ingredients) Silicone oil (Toray Dow Corning "SF8410") Organic gelling agent ("Gelall D" manufactured by New Japan Chemical Co., Ltd.) 1-Methyl-2-pyrrolidone (co-solvent)

[0181] Preparation of blood cell separation compositions A to H: The organic component having fluidity at 25° C., the inorganic fine powder, and other components were mixed in the blending ratios shown in Tables 1 and 2 to prepare compositions A to H for separating blood cells having thixotropy.

[0182] [Table 1]

[0183] [Table 2]

[0184] (red blood cell agglutinating agent) Dextran (weight average molecular weight 200,000) Hydroxyethyl starch-containing solution (Nipro's "HES40," an aqueous solution containing hydroxyethyl starch with a weight-average molecular weight of approximately 400,000 at a concentration of 6 w / v%) Polyethylene glycol (PEG 6K, Fujifilm Wako Pure Chemical Industries, Ltd.)

[0185] (osmolality adjuster) Sodium chloride glucose

[0186] (anticoagulant) Ethylenediaminetetraacetic acid dipotassium salt dihydrate (EDTA2K·2H2O)

[0187] (antioxidant) Sodium ascorbate

[0188] Example 1 A mixture was obtained by dissolving a red blood cell agglutinating agent, an osmotic pressure adjusting agent, and an anticoagulant in water. The types and amounts of the components in the mixture are shown in Table 3. The osmotic pressure of the mixture was adjusted to be isotonic with blood.

[0189] A polyethylene terephthalate bottomed tube (PET bottomed tube, blood collection container body) with a length of 100 mm and an inner diameter of the opening of 14 mm was prepared. 2.0 g of blood cell separation composition B was placed in the bottom of the blood collection container body. 1.0 mL of the resulting mixture was added to the surface of blood cell separation composition B. The pressure inside the blood collection container was reduced, and the container was sealed with a butyl rubber stopper. In this way, a blood collection container was produced. In the obtained blood collection container, the erythrocyte agglutinating agent, the osmotic pressure adjuster, and the anticoagulant were disposed on the surface of the blood cell separation composition in a liquid-dissolved state. The obtained blood collection container is a container for collecting 4 mL of blood.

[0190] (Examples 2 to 16 and Comparative Examples 3 and 5) Blood collection containers were prepared in the same manner as in Example 1, except that the type of blood cell separation composition and the amounts of the red blood cell agglutinating agent, anticoagulant, antioxidant, and osmotic pressure regulator were changed as shown in Tables 3 to 7. In the tables, the amount of hydroxyethyl starch-containing liquid is shown as the amount of hydroxyethyl starch-containing liquid (commercially available product) rather than the amount of hydroxyethyl starch contained. The osmotic pressure of the resulting mixed liquid was adjusted to be isotonic or hypertonic with blood.

[0191] (Comparative Example 1) The anticoagulant was dissolved in water to obtain a mixed solution.

[0192] A PET bottomed tube (blood collection container body) with a length of 100 mm and an inner diameter of the opening of 14 mm was prepared. 2.0 g of blood cell separation composition D was placed in the bottom of the blood collection container body. 30 mg of the resulting mixture was applied to the inner wall surface of the blood collection container body and allowed to dry. The pressure inside the blood collection container was reduced, and the container was sealed with a butyl rubber stopper. In this way, a blood collection container was produced. The resulting blood collection container was a container for collecting 4 mL of blood.

[0193] (Comparative Examples 2 and 4) A blood collection container was produced in the same manner as in Comparative Example 1, except that the type of composition for blood cell separation was changed as shown in Table 7.

[0194] (evaluation) (1) Specific gravity of the blood cell separation composition at 25°C A drop of the obtained composition for separating blood cells was dropped successively into saline solutions at 25° C. whose specific gravities had been adjusted stepwise at intervals of 0.002, and the specific gravities were measured by floating or sinking in the saline solutions.

[0195] (2) Osmotic pressure of the solution for measuring osmotic pressure 4 mL of physiological saline was added to the obtained blood collection container. After addition, the container was inverted to mix, dissolving the water-soluble components with the physiological saline, to obtain a solution for osmotic pressure measurement. The osmotic pressure of the obtained solution for osmotic pressure measurement was measured by the freezing point depression method using an osmometer ("OM-6060" manufactured by Arkray, Inc.).

[0196] (3) Red blood cell contamination rate and white blood cell recovery rate Blood samples from three individuals were collected and the following steps were carried out in order.

[0197] Blood collection process: 4 mL of blood was collected in the blood collection container body of the obtained blood collection container.

[0198] Centrifugation step: The blood collection vessel was centrifuged at 1500 G for 15 minutes.

[0199] After centrifugation, the plasma was located above the partition formed by the composition for blood cell separation. The plasma was stirred by pipetting to suspend the blood cells that had accumulated on the partition formed by the composition for blood cell separation, and then the blood cells were collected and used as a sample.

[0200] The collected samples were analyzed using a multi-parameter automated blood cell analyzer (Sysmex Corporation, "XE5000") to measure the number of white blood cells and red blood cells in the samples. The number of white blood cells and red blood cells in the prepared whole blood samples were also measured in the same manner. The number of white blood cells and red blood cells was the average value of the results obtained by evaluating the blood of three people.

[0201] The red blood cell contamination rate and white blood cell recovery rate were calculated using the following formulas.

[0202] Red blood cell contamination rate (%) = (number of red blood cells in the collected specimen (cells)) / (number of red blood cells in the whole blood sample (cells)) x 100

[0203] Leukocyte recovery rate (%) = (number of leukocytes in the collected specimen (cells)) / (number of leukocytes in the whole blood sample (cells)) x 100

[0204] [Criteria for determining red blood cell contamination rate] ○: Red blood cell contamination rate is 2% or less ×: Red blood cell contamination rate exceeds 2%

[0205] [Criteria for determining white blood cell recovery rate] ○: White blood cell recovery rate is 40% or more ×: White blood cell recovery rate is less than 40%

[0206] The compositions and results are shown in Tables 3 to 7 below.

[0207] [Table 3]

[0208] [Table 4]

[0209] [Table 5]

[0210] [Table 6]

[0211] [Table 7] [Explanation of symbols]

[0212] 1,1A…Blood collection container 2...Blood collection container body 2a...Inner wall surface 3...Composition for blood cell separation 4…Mixed liquid 4A…Mixed powder 5…Bung body

Claims

1. A leukocyte concentration and separation device to which a predetermined amount of a blood-derived sample is added, for concentrating and separating leukocytes present in the blood-derived sample, comprising: A container body; a blood cell separation material contained in the container body, the specific gravity of the blood cell separation material at 25°C is 1.082 or more and 1.090 or less, A leukocyte enrichment and separation device comprising the following configuration A1 and configuration B1. Configuration A1: A red blood cell agglutinating agent is contained in the container body. Configuration B1: The device is provided with an osmotic pressure adjusting agent contained within the container body, and when the water-soluble components contained within the container body are dissolved in a volume of physiological saline equal to the predetermined volume of the blood-derived sample to be added to the leukocyte concentration and separation device to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 320 mOsm / L or more and 360 mOsm / L or less.

2. A blood collection container into which a predetermined amount of blood is collected, A blood collection container body; a blood cell separation material contained in the blood collection container body, the specific gravity of the blood cell separation material at 25°C is 1.082 or more and 1.090 or less, A blood collection container comprising the following configuration A2 and configuration B2. Configuration A2: A red blood cell agglutinating agent is contained within the blood collection container body. Configuration B2: The blood collection container body is provided with an osmotic pressure adjuster, and when a solution for measuring osmotic pressure is obtained by dissolving the water-soluble component contained in the blood collection container body with a volume of physiological saline equal to the predetermined volume of blood to be collected in the blood collection container, the osmotic pressure of the solution for measuring osmotic pressure is 320 mOsm / L or more and 360 mOsm / L or less.

3. 3. The blood collection container according to claim 2, wherein the blood cell separation material is a blood cell separation composition having thixotropy.

4. 4. The blood collection container according to claim 2 or 3, wherein the red blood cell agglutinating agent is dextran, hydroxyethyl starch or polyethylene glycol.

5. 5. The blood collection container according to claim 4, wherein the weight-average molecular weight of the dextran is 50,000 or more.

6. 6. The blood collection container according to claim 2, wherein the osmotic pressure adjusting agent is sodium chloride or glucose.

7. The blood collection container of any one of claims 2 to 6, further comprising an anticoagulant contained within the blood collection container body.

8. The blood collection container of any one of claims 2 to 7, further comprising an antioxidant contained within the blood collection container body.

9. 9. The blood collection container of claim 8, wherein the antioxidant is ascorbic acid or an inorganic salt of ascorbic acid.

10. A method for separating leukocytes using the leukocyte concentration and separation device according to claim 1, adding a blood-derived sample to the leukocyte enrichment and separation device; and centrifuging the leukocyte concentration and separation device to which the blood-derived sample has been added.

11. A leukocyte concentration and separation system for collecting a predetermined amount of a blood-derived sample and concentrating and separating leukocytes present in the blood-derived sample, comprising: A blood cell separation material having a specific gravity of 1.082 or more and 1.090 or less at 25°C is provided, A leukocyte enrichment and separation system comprising the following configuration A3 and configuration B3. Configuration A3: Erythrocyte agglutinating agent is included. Configuration B3: An osmotic pressure adjusting agent is provided, and when a water-soluble component to be mixed with the blood-derived sample is dissolved in an amount of physiological saline equal to a predetermined amount of the blood-derived sample to obtain an osmotic pressure measurement solution, the osmotic pressure of the osmotic pressure measurement solution is 320 mOsm / L or more and 360 mOsm / L or less.

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