WO2015109644A1 - 细胞磁分选系统、分选装置和处理设备 - Google Patents

细胞磁分选系统、分选装置和处理设备 Download PDF

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Publication number
WO2015109644A1
WO2015109644A1 PCT/CN2014/073544 CN2014073544W WO2015109644A1 WO 2015109644 A1 WO2015109644 A1 WO 2015109644A1 CN 2014073544 W CN2014073544 W CN 2014073544W WO 2015109644 A1 WO2015109644 A1 WO 2015109644A1
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Prior art keywords
cell
outlet
solenoid
solution
shaped tube
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PCT/CN2014/073544
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English (en)
French (fr)
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张利峰
万书波
汪华
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张利峰
万书波
汪华
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Publication of WO2015109644A1 publication Critical patent/WO2015109644A1/zh
Priority to US15/214,440 priority Critical patent/US10053665B2/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/06—Magnetic means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/002—High gradient magnetic separation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/005—Pretreatment specially adapted for magnetic separation
    • B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/025—High gradient magnetic separators
    • B03C1/029—High gradient magnetic separators with circulating matrix or matrix elements
    • B03C1/03—High gradient magnetic separators with circulating matrix or matrix elements rotating, e.g. of the carousel type
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/025—High gradient magnetic separators
    • B03C1/031—Component parts; Auxiliary operations
    • B03C1/033—Component parts; Auxiliary operations characterised by the magnetic circuit
    • B03C1/0335—Component parts; Auxiliary operations characterised by the magnetic circuit using coils
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/23—Magnetic separation acting directly on the substance being separated with material carried by oscillating fields; with material carried by travelling fields, e.g. generated by stationary magnetic coils; Eddy-current separators, e.g. sliding ramp
    • B03C1/24—Magnetic separation acting directly on the substance being separated with material carried by oscillating fields; with material carried by travelling fields, e.g. generated by stationary magnetic coils; Eddy-current separators, e.g. sliding ramp with material carried by travelling fields
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/28—Magnetic plugs and dipsticks
    • B03C1/288—Magnetic plugs and dipsticks disposed at the outer circumference of a recipient
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/18—Magnetic separation whereby the particles are suspended in a liquid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/26—Details of magnetic or electrostatic separation for use in medical or biological applications
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1028—Sorting particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation

Definitions

  • Magnetic materials can specifically bind to target cells through biotechnology.
  • Using magnetically labeled target cells to magnetically classify highly specific target cells (target cells) is a critical step in cell medical and cell research activities.
  • the existing cell sorting schemes and cell sorting devices have disadvantages such as complicated operation, low sorting efficiency, low sorting purity, and insufficient control of the sorting environment. Summary of the invention
  • the object of the present invention is to overcome the deficiencies of the prior art and provide a cell magnetic sorting system, which has a simple structure, high efficiency and reliability, is suitable for continuous separation, can avoid environmental factors from contaminating the operation process, and can realize high quality cell division. selected.
  • Another object of the present invention is to provide a cell magnetic sorting apparatus for a cell magnetic sorting system. It is still another object of the present invention to provide a processing apparatus having the cell magnetic sorting system.
  • the present invention adopts the following technical solutions:
  • a cell magnetic sorting system comprising a cell solution source, a solution driving device, a cell continuous magnetic sorting device, a target cell collecting container, a raffinate collecting container and a control unit
  • the cell continuous magnetic sorting device comprises a rotating magnetic field generator a positive solenoid and an inverse solenoid that surround each other on the rotating magnetic field generator in a forward and reverse direction, one end of the positive solenoid is connected to the solution outlet of the cell solution source, and the other end is passed through ⁇ a tube is connected to one end of the counter solenoid, an inlet of the T-tube is in communication with an outlet of the positive solenoid, and a first outlet of the T-tube is connected to an inlet of the counter solenoid a second outlet connected to the inlet of the target cell collection container, the second outlet direction being consistent with the winding direction of the positive solenoid, the first outlet direction being opposite to the winding direction of the positive solenoid
  • the control unit connects the cell continuous magnetic sorting device and the solution driving device through
  • the inlet of the T-shaped tube and the initial portion of the first and second outlets are each provided with a flow stop switch. Also included is a device coupled to the second outlet of the tio tube for applying a negative pressure to the second outlet.
  • the solution driving device is driven by a solution pipe peristaltic extrusion method, comprising a main liquid pump disposed between the cell solution source and the continuous magnetic sorting device of the cell, and a continuous magnetic sorting device and a device disposed in the cell
  • the target cell collects the fluid pump between the containers.
  • the control unit includes a solution flow rate controller connected to the solution driving device through a control line and a magnetic field controller connected to the continuous magnetic sorting device of the cell, and the unfiltered cells are controlled by the solution flow rate controller and the magnetic field controller Magnetically labeled cells are reversely moved at the second exit of the tau tube to complete magnetic labeling cell sorting.
  • a processing apparatus having the cell magnetic sorting system.
  • a cell magnetic sorting device comprising a rotating magnetic field generator, a positive solenoid and an inverse solenoid wound on the rotating magnetic field generator in a forward and reverse direction, one end of the positive solenoid is connected to a solution outlet of the cell solution source, the other end being connected to one end of the counter solenoid through a stirrup tube, the inlet of the tau tube being in communication with an outlet of the positive solenoid, the stirrup tube
  • the first outlet is connected to the inlet of the counter solenoid
  • the second outlet is connected to the inlet of the target cell collection container
  • the second outlet direction is consistent with the winding direction of the positive solenoid
  • the first outlet direction is The winding directions of the positive solenoids are opposite.
  • the inlet of the stirrup tube and the initial portion of the first and second outlets are each provided with a flow stop switch.
  • the rotating magnetic field generator, the forward and reverse positive solenoids and the inverse solenoid design subtly realize the reverse spiral movement of the magnetically labeled cells in the opposite direction of the liquid flow, while the ⁇ -shaped tube design subtly subjects the magnetically labeled cells to the magnetic force.
  • the action moves against the solution flow direction and enters the target cell collection container, while the unlabeled cells migrate into the residue collection container in the direction of solution flow, thereby achieving continuous magnetic separation of the uninterrupted magnetic labeling cells, thereby reducing the sorting time and facilitating clinical Large-scale cell separation.
  • Magnetic labeling cell sorting is accomplished by controlling the reverse movement of unlabeled cells and magnetic labeling cells by the solution flow rate controller and the magnetic field controller, respectively.
  • the pipetting and sorting can be carried out at the same time, so that the cells can be quickly and effectively separated, and the cell sorting quality can be improved; the reverse solenoid design is beneficial to increase the number of cells sorted and improve Cell sorting purity; system can be cascaded to achieve continuous secondary sorting or continuous cell washing, reducing sorting.
  • the structure of the invention is simple, efficient and reliable, and the sorting is carried out in a closed pipeline, which can avoid pollution of the operation process by environmental factors, obtain high-volume and high-purity sorting cell products, and facilitate sorting of clinical grade cells. product.
  • control unit can control the rinsing and recovery of the target cells through the control line, and the cell separation, rinsing and recovery canning process is completed once.
  • FIG. 2 is a schematic structural view of an embodiment of a continuous magnetic sorting device for cells according to the present invention
  • Figure 3 is a plan view of the continuous magnetic sorting apparatus of the cell shown in Figure 2.
  • the cell magnetic sorting system comprises a cell solution source-K cell continuous magnetic sorting device 2, a raffinate collection container 3, a target cell collection container 4, a main liquid pump 5, and a slave
  • the cell continuous magnetic sorting device 2 includes a rotating magnetic field generator 13, positive and negative solenoids 12, 12' which are wound on the rotating magnetic field generator 13 in forward and reverse directions, and are connected to the positive and negative solenoids 12, 12 ! T-tube 11 between.
  • the cell solution source 1 is connected to the solution inlet of the cell continuous magnetic sorting device 2 via the main liquid pump 5, and the residual liquid outlet of the cell continuous magnetic sorting device 2 is connected to the raffinate collection container 3, and the T-shaped tube of the cell continuous magnetic sorting device 2
  • the second outlet of 1 1 is connected to the target cell collection container 4 from the liquid pump 6 .
  • the control line of the solution flow rate controller 7 is connected to the main liquid pump 5 and the liquid accumulating pump 6.
  • the control line of the magnetic field controller 8 is connected to the cell continuous magnetic sorting device 2.
  • the solution flow rate controller 7 controls the main liquid pump 5 to extract the magnetically labeled cell solution from the cell solution source 1 and inject it into the cell continuous magnetic sorting device 2, and the control circuit of the magnetic field controller 8 controls the continuous magnetic sorting device 2
  • the rotating magnetic field generator 13 generates a rotating magnetic field, applies a magnetic rotating force to the magnetically labeled cells flowing into the solenoid, and moves the magnetically labeled cells in a reverse spiral opposite to the direction in which the liquid flows, thereby achieving target cell sorting.
  • the solution flow rate controller 7 drives the liquid crystal pump 6 to move the sorted target cell solution into the target cell collection container 4, and the remaining residual solution is introduced into the residue collection container 3.
  • the main liquid pump 5 provides a driving force for injecting the cells from the cell solution source 1 and injecting the cells into the continuous magnetic sorting device 2, and also provides a driving force for injecting the cell solution into the target cell collecting container 4 and the raffinate collecting container 3.
  • the driving force provided by the liquid pump 6 compensates for the flow of the target cell solution after separation Speed to improve cell separation efficiency.
  • one end of the positive solenoid 12 serves as a liquid inlet 9 connected to the source 1 of the cell solution, and the other end of the positive solenoid 12
  • the T-shaped tube 1 1 is connected to one end of the counter solenoid 12', and the other end of the counter solenoid 12' serves as the liquid outlet 10 connected to the raffinate collecting container 3, and the opposite ends of the T-shaped tube 1 1
  • the positive solenoid 12 and the counter solenoid 12' are respectively connected, and the first outlet of the manifold 11 is connected to the inlet of the counter solenoid 12', and the first outlet direction is opposite to the winding direction of the positive solenoid 12.
  • the second outlet direction of the stirrup tube 11 coincides with the winding direction of the positive solenoid tube 2, and the second outlet of the sickle tube 1] is a sorting port that communicates with the target cell collection container 4.
  • the rotating magnetic field component 13 absorbs and moves the target cell in the rotating magnetic field generated in the positive and negative solenoids 12, 12', in the positive phase of the solenoid 12 segment, that is, the solenoid.
  • the magnetically labeled target cells are accelerated by the magnetic field to move toward the sorting port of the fistula 1 1 , and in the reverse phase of the reverse solenoid 12 ', ie, the solenoid, the magnetically labeled target cells are reversed by the magnetic field.
  • the liquid flows toward the sorting port of the sickle tube 11, so that the magnetically labeled target cells are concentrated in the sorting port of the sickle tube 11 and separated and discharged to the target cell collection container 4.
  • reference numeral 18 denotes a graph showing the state of stress of magnetically labeled cells and unlabeled cells at various stages of the continuous magnetic sorting apparatus 2 of the cells.
  • a black solid circle indicates magnetically labeled cells
  • a hollow circle indicates unlabeled cells
  • a thinner arrow indicates liquid flow force
  • a thicker arrow indicates magnetic force.
  • the cell solution is input from the sputum port 9, wherein the magnetically labeled target cells rapidly reach the scorpion tube under the action of the positive solenoid 12 and the rotating magnetic field 5, and the interaction between the counter solenoid 12' and the rotating magnetic field 5
  • the magnetically labeled cells create resistance that prevents them from continuing to flow along the counter-rotor, whereby the magnetically labeled target cells are enriched at the fistula 11 and discharged from the sorting port.
  • the target cells are rinsed and canned; an appropriate negative pressure can be applied at the sorting port of the stirrup tube 11 to extract the enriched target cells.
  • the unlabeled magnetic cells have no magnetic force due to the magnetic field, and there is no phenomenon that the positive solenoid 12 is accelerated, the reverse solenoid 12' is blocked, and the scorpion tube is enriched.
  • the solenoid 12' is led out of the liquid outlet 10.
  • the control circuit of the magnetic field controller 8 can also be connected to the solution flow rate controller 7, and the solution flow rate controller 7 and the magnetic field controller 8 can coordinately drive the cell continuous magnetic sorting device 2 to achieve efficient, high quality cell sorting effects.
  • Other embodiments are directed to a processing device that can be a variety of devices for cell medical or cellular research, which can have the cell magnetic sorting system of any of the above embodiments.

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Abstract

本发明提供了一种细胞磁分选系统，其包括细胞溶液源、溶液驱动装置、细胞连续磁分选装置、目标细胞收集容器、残液收集容器和控制单元。所述细胞连续磁分选装置包括旋转磁场发生器、互为正反向地环绕在旋转磁场发生器上的正螺线管和逆螺线管，正螺线管的一端连通到细胞溶液源，另一端通过T形管与逆螺线管的一端连接，T形管的入口与正螺线管的出口连通，T形管的第一出口连通到逆螺线管的入口，第二出口连通到目标细胞收集容器的入口，第二出口方向与正螺线管绕线方向一致，第一出口方向与正螺线管绕线方向相反。本发明还提供了一种细胞磁分选装置和一种处理设备。

Description

磁性材料通过生物技术可以与靶细胞特异性结合， 利用磁标记的靶细 胞的磁性， 用电磁方法分选出高度特异性的靶细胞（目标细胞），是细胞医 疗和细胞科研活动的关键性步骤。 现有的细胞分选方案和细胞分选装置存 在着操作较复杂、分选效率低、分选纯度不高和分选环境控制不足等弊端。 发明内容
本发明的目的就是为了克服现有技术的不足， 提供一种细胞磁分选系 统， 其结构简单， 高效可靠， 适合连续分离、 能避免环境因素对操作过程 的污染， 可实现高质量的细胞分选。
本发明的另一目的是提供用于细胞磁分选系统的细胞磁分选装置。 本发明的又一目的是提供具有该细胞磁分选系统的处理设备。
为实现上述目的， 本发明采用以下技术方案：
一种细胞磁分选系统， 包括细胞溶液源、 溶液驱动装置、 细胞连续磁 分选装置、 目标细胞收集容器、 残液收集容器和控制单元， 所述细胞连续 磁分选装置包括旋转磁场发生器、 互为正反向地环绕在所述旋转磁场发生 器上的正螺线管和逆螺线管， 所述正螺线管的一端连通到所述细胞溶液源 的溶液出口， 另一端通过 τ形管与所述逆螺线管的一端连接， 所述 T形管 的入口与所述正螺线管的出口连通， 所述 T形管的第一出口连通到所述逆 螺线管的入口， 第二出口连通到所述目标细胞收集容器的入口， 第二出口 方向与所述正螺线管的绕线方向一致， 第一出口方向与所述正螺线管的绕 线方向相反， 所述控制单元通过控制线路连接所述细胞连续磁分选装置和 所述溶液驱动装置， 用于控制所述细胞连续磁分选装置产生磁作 ^力， 并 控制所述溶液驱动装置将所述细胞溶液源中的溶液驱动， 经所述 τ 形管， 使未标记细胞和磁标记细胞分别流入到所述残液收集容器和所述目标细 胞收集容器。
所述 T形管的入口和第一、 第二出口的起始部分均设置有液流停止开 关。 还包括与所述 τ形管的第二出口相连用于向所述第二出口处施加负压 的装置。
所述溶液驱动装置采用溶液管道蠕动挤压方式驱动， 包括设置在所述 细胞溶液源和所述细胞连续磁分选装置之间的主液泵和设置在所述细胞 连续磁分选装置和所述目标细胞收集容器之间的从液泵。
所述控制单元包括通过控制线路连接所述溶液驱动装置的溶液流速 控制器和连接所述细胞连续磁分选装置的磁场控制器， 通过所述溶液流速 控制器和磁场控制器控制未标记细胞和磁标记细胞在所述 τ形管的第二出 口处反向运动完成磁标记细胞分选。
一种处理设备， 具有所述的细胞磁分选系统。
一种细胞磁分选装置， 包括旋转磁场发生器、 互为正反向地环绕在所 述旋转磁场发生器上的正螺线管和逆螺线管， 所述正螺线管的一端连通到 所述细胞溶液源的溶液出口， 另一端通过 Τ形管与所述逆螺线管的一端连 接， 所述 τ形管的入口与所述正螺线管的出口连通， 所述 Τ形管的第一出 口连通到所述逆螺线管的入口， 第二出口连通到所述目标细胞收集容器的 入口， 第二出口方向与所述正螺线管的绕线方向一致， 第一出口方向与所 述正螺线管的绕线方向相反。
所述 Τ形管的入口和第一、 第二出口的起始部分均设置有液流停止开 关。
还包括与所述 Τ形管的第二出口相连用于向所述第二出口处施加负压 的装置。
本发明有益的技术效果在于：
旋转磁场发生器、 互为正反向的正螺线管和逆螺线管设计巧妙地实现 磁标记细胞与液体流动方向相反的逆向螺旋移动， 同时 τ形管设计巧妙地 使磁标记细胞受到磁力作用逆溶液流动方向移动迸入目标细胞收集容器， 而未标记的细胞顺溶液流动方向移行进入残液收集容器， 实现不间断的磁 标记细胞连续磁分选， 从而减少分选时间， 有利于临床级别大规模的细胞 分离。 通过所述溶液流速控制器和磁场控制器分别控制未标记细胞和磁标 记细胞反向运动方式完成磁标记细胞分选。
通过控制电磁流体和溶液流体的反向运动， 使移液和分选同时迸行， 实现细胞快速有效分离， 可提高细胞分选质量； 逆向螺线管设计， 有利于 增加细胞分选数量， 提高细胞分选纯度； 系统可级联， 从而实现连续二次 分选或连续细胞清洗， 减少分选 间。 本发明结构筒单， 高效可靠， 分选处在密闭的管道中进行， 能避免环 境因素对操作过程的污染， 可获得高数量和高纯度的分选细胞产品， 有利 于分选临床级别的细胞制品。
进一步地， 通过在 T形管入口和出口的起始部分设置液流停止开关， 方便控制单元通过控制线路控制目标细胞的漂洗和回收， 实现细胞分离、 漂洗和回收罐装工艺一次完成。
¾ 、图 1为本发明细胞磁分选系统实施例的结构示意图；
图 2为本发明细胞连续磁分选装置实施例的结构示意图；
图 3为图 2所示的细胞连续磁分选装置的俯视图。
具体实施方式
以下通过实施例结合附图对本发明迸行进一步的详细说明。
参阅图 1至图 3， 在一些实施例里， 细胞磁分选系统包括细胞溶液源 -K 细胞连续磁分选装置 2、 残液收集容器 3、 目标细胞收集容器 4、 主液 泵 5、 从液泵 6、 溶液流速控制器 7以及磁场控制器 8。 细胞连续磁分选装 置 2包括旋转磁场发生器 13、 互为正反向地环绕在旋转磁场发生器 13上 的正、 逆螺线管 12、 12 ' 和连接在正、 逆螺线管 12、 12 ! 之间的 T形管 11。 细胞溶液源 1经过主液泵 5连接细胞连续磁分选装置 2的溶液入口， 细胞连续磁分选装置 2的残液出口连接残液收集容器 3， 细胞连续磁分选 装置 2的 T形管 1 1的第二出口即分选口经过从液泵 6连接到目标细胞收集 容器 4。溶液流速控制器 7的控制线路连接主液泵 5和认液泵 6。磁场控制 器 8的控制线路连接细胞连续磁分选装置 2。
使用时， 溶液流速控制器 7控制主液泵 5从细胞溶液源 1吸取经磁标 记的细胞溶液并注入细胞连续磁分选装置 2， 磁场控制器 8的控制线路控 制连续磁分选装置 2的旋转磁场发生器 13产生旋转磁场，施加磁旋转作用 力到流入螺线管中的磁标记的细胞上， 使磁标记细胞与液体流动方向相反 的逆向螺旋移动， 实现目标细胞分选。 溶液流速控制器 7驱动从液泵 6将 分选后的目标细胞溶液移入目标细胞收集容器 4， 剩余残余溶液导入残液 收集容器 3。
主液泵 5除了提供从细胞溶液源 1取液并注入细胞连续磁分选装置 2 的驱动力， 还提供推动细胞溶液分离后注入目标细胞收集容器 4和残液收 集容器 3的驱动力。 从液泵 6提供的驱动力可补偿分离后目标细胞溶液流 速以提高细胞分离效率。 通过增设从液泵 6， 控制其配合主液泵 5协同工 作， 可提高目标细胞的分选速度和分选质量。
参阅图 2和图 3 , 在一些实施例里， 细胞连续磁分选装置 2中， 正螺 线管 12的一端作为连通到细胞溶液源 1的进液口 9， 正螺线管 12的另一 端通过 T形管 1 1与逆螺线管 12 ' 的一端连接， 逆螺线管 12 ' 的另一端作 为连通到残液收集容器 3的出液口 10，T形管 1 1的两个对端分别连接正螺 线管 12和逆螺线管 12 ' ， Τ形管 11的第一出口连通到逆螺线管 12 ' 的入 口，第一出口方向与正螺线管 12的绕线方向相反， Τ形管 11的第二出口方 向与正螺线管 ] 2的绕线方向一致， Τ形管 1 ] 的第二出口为连通到目标细 胞收集容器 4的分选口。 工作过程中， 在磁场控制器控制下， 旋转磁场部 件 13在正逆螺线管 12、 12 ' 中产生的旋转磁场吸 并移动目标细胞， 在 正螺线管 12段即螺线管正向阶段,经过磁标记的目标细胞受磁场作用加速 向 Τ形管 1 1的分选口移动， 而在逆螺线管 12 ' 段即螺线管反向阶段， 经 过磁标记的目标细胞受磁场作用逆液体流向向 Τ形管 11的分选口移动，从 而,经过磁标记的目标细胞在 Τ形管 1 1的分选口富集并分离排出至目标细 胞收集容器 4。
参阅图 2，标号 18代表的图形显示了磁标记细胞和未标记细胞在细胞 连续磁分选装置 2各阶段受力状态。 图中， 黑色实心圆表示磁标记细胞， 空心圆表示未标记细胞， 较细的箭头表示液体流动力， 较粗的箭头表示磁 作用力。 细胞溶液从迸液口 9输入， 其中经磁标记的目标细胞在正螺线管 12和旋转磁场 5共同作用下快速到达 Τ形管处，逆螺线管 12 ' 和旋转磁场 5的共同作用对磁标记细胞产生阻力， 阻止其继续沿逆螺线管流出， 由此， 经磁标记的目标细胞会富集在 Τ形管 11处并从分选口排出。当 Τ形管第二 出口起始部开关关闭时， 可实行双向螺线管道冲洗； 当 Τ形管第一出口起 始部开关关闭时， 可实行 Τ形管第二出口管道和细胞收集容器的目标细胞 漂洗和罐装;可在 Τ形管 11的分选口处施加适当的负压以便将富集的目标 细胞引出。 另外， 未标记磁的细胞由于磁场对其无磁力作用， 不会出现被 正螺线管 12加速、 被逆螺线管 12 ' 阻止、 在 Τ形管处富集的现象， 故会 顺利通过逆螺线管 12 ' 从出液口 10导出。
磁场控制器 8的控制线路也可以连接溶液流速控制器 7， 溶液流速控 制器 7和磁场控制器 8可协调驱动细胞连续磁分选装置 2， 达到高效、 高 质量的细胞分选效果。 另一些实施例中是关于一种处理设备， 该处理设备可以是用于细胞医 疗或细胞科研的各种设备，其可具有上述任一种实施例的细胞磁分选系统。
以上内容是结合具体的优选实施方式对本发明所作的进一步详细说 明， 不能认定本发明的具体实施只局限于这些说明。 例如， 所属技术领域 的普通技术人员能够理解， 提供液体流动力的装置除了采用液泵之外， 也 可以釆用其他任何能够驱动液流的设备。 对于本发明所属技术领域的普通 技术人员来说， 在不脱离本发明构思的前提下， 还可以做出若干筒单推演 或替换， 都应当视为属于本发明的保护范围。

Claims

权 利 要 求 书
1, 一种细胞磁分选系统， 其特征在于，包括细胞溶液源、 溶液驱动装 置、细胞连续磁分选装置、 目标细胞收集容器、残液收集容器和控制单元， 所述细胞连续磁分选装置包括旋转磁场发生器、 互为正反向地环绕在所述 旋转磁场发生器上的正螺线管和逆螺线管， 所述正螺线管的一端连通到所 述细胞溶液源的溶液出口，另一端通过 τ形管与所述逆螺线管的一端连接， 所述 τ形管的入口与所述正螺线管的出口连通， 所述 T形管的第一出口连 通到所述逆螺线管的入口，第二出口连通到所述目标细胞收集容器的入口， 第二出口方 与所述正螺线管的绕线方向一致， 第一出口方 与所述正螺 线管的绕线方向相反， 所述控制单元通过控制线路连接所述细胞连续磁分 选装置和所述溶液驱动装置， 用于控制所述细胞连续磁分选装置产生磁作 用力， 并控制所述溶液驱动装置将所述细胞溶液源中的溶液驱动， 经所述 τ 形管，使未标记细胞和磁标记细胞分别流入到所述残液收集容器和所述 目标细胞收集容器。
2, 如权利要求 1所述的细胞磁分选系统， 其特征在于,所述 T形管的 入口和第一、 第二出口的起始部分均设置有液流停止开关。
3, 如权利要求 1所述的细胞磁分选系统， 其特征在于，还包括与所述 T形管的第二出口相连用于向所述第二出口处施加负压的装置。
4, 如权利要求 1所述的细胞磁分选系统， 其特征在于，所述溶液驱动 装置采用溶液管道蠕动挤压方式驱动， 包括设置在所述细胞溶液源和所述 细胞连续磁分选装置之间的主液泵和设置在所述细胞连续磁分选装置和 所述目标细胞收集容器之间的从液泵。
5, 如权利要求]至 4任一项所述的细胞磁分选系统，其特征在于， 所 述控制单元包括通过控制线路连接所述溶液驱动装置的溶液流速控制器 和连接所述细胞连续磁分选装置的磁场控制器， 通过所述溶液流速控制器 和磁场控制器控制未标记细胞和磁标记细胞在所述 T形管的第二出口处反 向运动完成磁标记细胞分选。
6, —种处理设备， 其特征在于， 具有如权利要求 1至 5任一项所述 的细胞磁分选系统。
7, 一种细胞磁分选装置， 其特征在于，包括旋转磁场发生器、 互为正 反向地环绕在所述旋转磁场发生器上的正螺线管和逆螺线管， 所述正螺线 管的一端连通到所述细胞溶液源的溶液出口， 另一端通过 τ形管与所述逆 螺线管的一端连接， 所述 τ形管的入口与所述正螺线管的出口连通， 所述 τ 形管的第一出口连通到所述逆螺线管的入口， 第二出口连通到所述目标 细胞收集容器的入口， 第二出口方向与所述正螺线管的绕线方向一致， 第 一出口方向与所述正螺线管的绕线方向相反。
8. 如权利要求 7所述的细胞磁分选装置， 其特征在于，所述 T形管的 入口和第一、 第二出口的起始部分均设置有液流停止开关。
9, 如权利要求 7所述的细胞磁分选装置， 其特征在于，还包括与所述 T形管的第二出口相连用于向所述第二出口处施加负压的装置。
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