CN211188500U

CN211188500U - Cell separation and feedback device

Info

Publication number: CN211188500U
Application number: CN201920925371.9U
Authority: CN
Inventors: 强斌; 廉云飞; 陆晓; 黄静; 韩静妮
Original assignee: Jiangsu Tuohong Kangheng Pharmaceutical Co ltd
Current assignee: Jiangsu Tuohong Kangheng Pharmaceutical Co ltd
Priority date: 2019-06-19
Filing date: 2019-06-19
Publication date: 2020-08-07
Anticipated expiration: 2029-06-19

Abstract

The utility model relates to a cell separation and feedback device, including the separator tube that is the tube-shape, the separator tube endotheca is equipped with the movable assembly with separator tube sliding connection, the movable assembly includes a piston axle, the one end fixedly connected with piston of piston axle, the piston includes the first surface and the second surface that are parallel to each other, the axial setting of first surface and second surface perpendicular to piston axle, the internal surface of first surface and separator tube forms one and is used for the holding to treat the cavity of separating cell liquid, treat including the target cell in the separating cell liquid, still the cover is equipped with the annular magnetic force separating unit with piston axle sliding connection on the piston axle, annular magnetic force separating unit be located be close to one side on piston second surface and with the second surface parallels, the separator tube still is provided with first pipeline and the second pipeline with the cavity intercommunication, annular magnetic force separating unit is connected with the second surface contact and relies on magnetic force will combine the magnetic substance of target cell to adsorb on the first surface.

Description

Cell separation and feedback device

Technical Field

The utility model relates to the field of medical equipment, especially, relate to a cell separation and feedback device.

Background

Cells often exist in tissues or blood, and the precise research on a certain specific cell requires the separation and purification of the specific cell from different types of tissues and blood, and CN207908244U discloses a fully automatic blood cell separation and purification device, which has a complex structure and high cost, and cannot realize the instant separation and feedback of the cell.

In clinical practice, hematopoietic stem cell transplantation has become an important therapeutic approach, and is applied to the treatment of various serious diseases. In addition to bone marrow, there are some hematopoietic stem cells in normal human peripheral blood, and if the hematopoietic stem cells in bone marrow are "mobilized" by drugs and then released into peripheral blood, one can directly collect hematopoietic stem cells by peripheral blood sorting, and the mobilizing agent generally increases the number of CD34 positive cells in peripheral blood by 20-30 times. This is also currently the most commonly used method in hematopoietic stem cell transplantation. In addition, in the delivery of a term fetus, hematopoietic stem cells are also transferred from a fetal liver to bone marrow, so that a certain amount of hematopoietic stem cells are contained in cord blood, and 40 to 200 ten thousand cells containing CD34+ are contained in 100 ml of cord blood.

Diabetic foot is one of the serious chronic complications that cause disability and death in diabetic patients. The world health organization defines a diabetic foot as a diabetic patient with a combination of neuropathy and various peripheral vascular lesions of varying degrees that result in lower limb infections, ulceration and/or destruction of deep tissues. The traditional treatment means of the diabetic foot comprise conservative treatments such as medicines, vascular bypass, interventional operation and the like, the effect is poor, and in clinic, at least 30-40 percent of diabetic foot patients have to receive amputation operation due to poor microcirculation of lower limbs and complete occlusion of small blood vessels, so that the surgical vascular reconstruction operation cannot be performed. Therefore, it is a necessary trend to open new therapeutic approaches.

The stem cell transplantation for treating diabetic foot is a new method appearing in recent years, is rapidly developed and is applied to clinic. According to different sources of stem cells, the current clinical methods for treating diabetic foot by autologous stem cell transplantation mainly include autologous bone marrow stem cell transplantation and autologous peripheral blood stem cell transplantation. The bone marrow stem cell transplantation is an early developed method, and the autologous peripheral blood stem cell transplantation has the advantages of convenient collection and little pain for patients.

According to the principle that HSC can be differentiated into vascular endothelial cells and smooth muscle cells under the induction action, a large number of angiogenesis promoting factors are secreted, and the formation of new blood vessels is promoted, the autologous peripheral blood CD34+ HSC of a patient is separated and transplanted to an ischemic lower limb part, so that the autologous peripheral blood CD34+ hematopoietic stem cells can be gradually differentiated to form new blood capillaries, the blood flow of the lower limb is improved and recovered, and the new blood vessels can grow after 3-6 months of recovery, so that the purpose of treating diabetic feet is achieved. Because the HSC is taken from the self, no immune rejection reaction exists, and the safety of clinical application is ensured.

Autologous HSC transplantation, as a new technique for treating diabetic foot, has been widely used and successful at home and abroad, and can relieve amputation or reduce amputation level in some patients and improve the quality of life of the patients. The current methods for obtaining autologous HSCs all use a mononuclear cell separator. The mononuclear cell separator is expensive and cannot be popularized, so that the use of a diabetic foot patient is greatly limited. Therefore, it is important to develop a technique for easily isolating autologous HSCs, and to popularize this method.

SUMMERY OF THE UTILITY MODEL

In order to solve the above technical problem, the present invention provides a cell separating and feedback device, which has a simple structure and a low cost, and can realize the instant separation and feedback of cells.

The utility model discloses a cell separation and feedback device, including the separator tube that is the tube-shape, the separator tube endotheca is equipped with the movable assembly with separator tube sliding connection, the movable assembly includes a piston axle, the one end fixedly connected with piston of piston axle, the piston includes first surface and the second surface that is parallel to each other, first surface and second surface perpendicular to the axial setting of piston axle, form a cavity that is used for holding the cell liquid that waits to separate between the internal surface of first surface and the separator tube, including the purpose cell in the cell liquid that waits to separate, still the cover is equipped with the annular magnetic force separation unit with piston axle sliding connection on the piston axle, annular magnetic force separation unit is located the one side that is close to the piston second surface and parallel with the second surface of piston, the separator tube still be provided with the first pipeline and the second pipeline of cavity intercommunication, the first pipeline is used for inputting the cell fluid to be separated, the second pipeline is used for inputting the magnetic substance combined with the target cells, when the annular magnetic separation unit is connected with the second surface in a contact mode, the magnetic substance combined with the target cells is adsorbed on the first surface, then the piston slides towards the side far away from the second surface relative to the separation pipe, and the separated cell fluid is extruded out of the separation pipe.

Further, the separated cell liquid is output from the first pipeline.

Further, after the separated cell sap is extruded from the separation tube, the annular magnetic separation unit is separated from the second surface, and the magnetic substance bound to the target cell falls off from the first surface, so that the separation of the target cell is realized.

Furthermore, an opening is formed in one end of the separation pipe, a blocking piece used for blocking the end is arranged at the other end of the separation pipe, and the piston shaft and the piston are sleeved in the opening.

Furthermore, the baffle plate is provided with a first through hole and a second through hole, and the first pipeline and the second pipeline are respectively communicated with the first through hole and the second through hole.

Furthermore, one end of the first pipeline, which is far away from the separation pipe, is connected with a collection device and/or a liquid outlet pipe for collecting cell sap to be separated.

Furthermore, one end of the second pipeline, which is far away from the separation pipe, is connected with a liquid inlet pipe and/or a bottle stopper puncture outfit.

Furthermore, a control clamp for controlling the flow rate is arranged on the first pipeline and/or the second pipeline.

Further, one end of the first pipeline and/or the second pipeline, which is far away from the separation pipe, is connected with a luer. The first tubing and/or the second tubing may be removably connected to other medical products via a luer.

Further, the magnetic substance is an antibody-coated magnetic bead. The antibody is used for recognizing the target cells, and the magnetic beads are used for being adsorbed by the annular magnetic separation unit.

Further, the first pipeline and the second pipeline are parallel to the axial direction of the separation pipe.

Further, one end of the piston shaft, which is far away from the piston, is fixedly connected with a piston handle.

Further, the diameter of the annular magnetic separation unit is not larger than the inner diameter of the separation tube.

Further, the target cells are autologous peripheral blood stem cells.

Borrow by above-mentioned scheme, the utility model discloses at least, have following advantage:

the utility model discloses can realize the instant separation to the cell, make things convenient for clinical application, through the setting of piston and annular magnetic separation unit and the change of position relation, can realize the collection feedback of autologous cell, avoid the risk of xenogenesis infusion, simultaneously in the treatment, can separate immediately to the cell, instant feedback has shortened single treatment time, has reduced treatment cost.

The above description is only an overview of the technical solution of the present invention, and in order to make the technical means of the present invention clearer and can be implemented according to the content of the description, the following detailed description is made with reference to the preferred embodiments of the present invention and accompanying drawings.

Drawings

FIG. 1 is a schematic structural view of a separation tube according to the present invention;

FIG. 2 is a schematic structural view of the movable assembly of the present invention;

FIG. 3 is a schematic structural view of the annular magnetic separation unit of the present invention;

fig. 4 is a schematic structural diagram of the present invention in an operating state;

fig. 5 is a schematic structural view in another working state of the present invention;

description of reference numerals:

101-a separation tube; 102-a first interface; 103-a second interface; 104-a piston; 1041-a first surface; 1042 — a second surface; 105-an annular magnetic separation unit; 106 — a first conduit; 107-second line; 108-a piston shaft; 109-piston shank; 110-magnetic beads to which cells of interest are bound.

Detailed Description

The following detailed description of the embodiments of the present invention is provided with reference to the accompanying drawings and examples. The following examples are intended to illustrate the invention, but are not intended to limit the scope of the invention.

Example 1

The embodiment provides a clinical grade autologous peripheral blood stem cell collection and return device, which can realize clinical grade stem cell collection, instant separation and return transfusion, and hereinafter, the term "mixed liquid" is mixed with other components and target cells, and the mixed liquid realizes cell separation in a separation tube.

The cell separation device provided by the utility model can be used for the separation of stem cells and other blood components. In the examples, "magnetic beads" used for separating stem cells refer to magnetic materials used for sorting cells, the surface of the magnetic beads carrying antibodies that recognize the cells of interest, such as those derived from PBMA containing CD3⁺/CD28⁺Magnetic microbeads of antibodies isolate T cells.

The utility model discloses in, piston and annular magnetic separation unit all rely on the motion of manual realization relative separation pipe.

Referring to fig. 1-3, a cell separation and feedback device according to a preferred embodiment of the present invention includes a cylindrical separation tube 101, wherein one end of the separation tube 101 is provided with an opening, and the other end is provided with a blocking piece for blocking. The separating tube 101 is sleeved with a movable assembly which is connected with the separating tube in a sliding manner, and the movable assembly comprises a piston shaft 108, a piston 104 and a piston handle 109. The piston shaft 108 and the piston 104 are located inside the separation tube 101, and the piston handle 109 is located outside the separation tube 101. The piston 104 and the piston handle 109 are fixedly connected to both ends of the piston shaft 108, respectively. The piston 104 includes a first surface 1041 and a second surface 1042 which are parallel to each other, the first surface 1041 and the second surface 1042 are perpendicular to the axial direction of the piston shaft 108, and when the piston 104 is not in contact with the baffle, a cavity for containing the mixed liquid is formed between the first surface 1041 and the inner surface of the separation tube 101.

The separation pipe 101 is further provided with a first pipeline 106 and a second pipeline 107 communicated with the cavity, and the first pipeline 106 and the second pipeline 107 are parallel to the axial direction of the separation pipe 101. The first pipeline 106 is used for inputting mixed liquid, and the second pipeline 107 is used for inputting magnetic beads coated by antibodies. A first through hole and a second through hole are formed in the baffle plate of the separation pipe 101, the first pipeline 106 is communicated with the cavity body sequentially through the first interface 102 and the first through hole, and the second pipeline 107 is communicated with the cavity body sequentially through the second interface 103 and the second through hole.

One end of the first pipeline 106, which is far away from the separation tube 101, is connected with a blood taking needle, and blood taking and blood return transfusion can be performed. One end of the second pipeline 107 far away from the separation tube 101 is connected with a bottle stopper puncture outfit for injecting and adding magnetic beads or other medicine components into the separation tube 101.

The last control that can also be equipped with of first pipeline 106 and second pipeline 107 presss from both sides for the flow of the mixed liquid of control, all control that can adjust liquid flow press from both sides all can be applied to the utility model discloses. The ends of the first and

second lines

106, 107 remote from the separation tube 101 may also be connected to a luer. The first tubing 106 and/or the second tubing 107 may be removably connected to other medical products via a luer.

The piston shaft 108 is further sleeved with an annular magnetic separation unit 105 slidably connected with the piston shaft 108, the annular magnetic separation unit 105 is located at a side close to the second surface 1042 of the piston 104 and is parallel to the second surface 1042, and the annular magnetic separation unit 105 can freely move along the piston shaft 108 relative to the piston 104. The annular magnetic separation unit 105 is in the shape of a disc, which may have a diameter smaller than the inner diameter of the separation tube 101. When the annular magnetic separation unit 105 is in contact with the second surface 1042, for example, the separation tube 101 is vertically placed with its axial direction perpendicular to the ground, and then the annular magnetic separation unit 105 contacts with the second surface 1042 by gravity, after a certain time of contact, the antibodies on the magnetic beads are sufficiently bound to the target cells, the magnetic beads 110 bound to the target cells are adsorbed on the first surface 1041, the second tube 107 is closed, it is ensured that the annular magnetic separation unit 105 contacts with the second surface 1042 and pushes the piston 104 towards the side away from the second surface 1042 relative to the separation tube 101, so that the separated cellular fluid is squeezed out from the separation tube 101, and the separated cellular fluid is output from the first tube 106. After the separated cell fluid is extruded from the separation tube 101, the annular magnetic separation unit 105 is separated from the second surface 1042, and the magnetic beads 110 bound with the target cells are separated from the first surface 1041, so that the target cells are separated.

Use the target cell to be CD34 cell as the example, the utility model discloses at the during operation, at first adopt blood through the blood taking needle, the blood of collection gets into separator tube 101 through first pipeline 106. The container containing the antibody-coated magnetic beads is then pierced by the stopper piercer and the antibody-coated magnetic beads enter the separation tube 101 through the second conduit 107. The blood and the antibody-coated magnetic beads are mixed in the separation tube 101 to form a mixed solution. In the above process, the first surface 1041 of the piston 104 is spaced from the baffle of the separation tube 101, so that the separation tube 101 has enough space to accommodate the mixed liquid. Referring to fig. 4, the separation tube 101 is vertically disposed with its axial direction perpendicular to the ground, the first surface 1041 is located below the second surface 1042, the annular magnetic separation unit 105 contacts the second surface 1042 by gravity, after a certain time, the antibodies on the magnetic beads are sufficiently combined with the CD34 cells, the magnetic beads with CD34 cells are adsorbed on the first surface 1041, the second tube 107 is closed, the piston 104 is manually pushed toward the side away from the second surface 1042, and the separated cellular fluid is output from the first tube 106. In the process, since the diameter of the annular magnetic separation unit 105 is smaller than the inner diameter of the separation tube 101, when the separation tube 101 is vertically placed, the annular magnetic separation unit 105 can slide downwards along the piston shaft 108 along with the piston 104 by virtue of gravity, so as to ensure that the annular magnetic separation unit 105 is always in contact with the second surface 1042 of the piston 104, and thus, the magnetic beads combined with the CD34 cells are always adsorbed on the first surface 1041 and cannot be output from the separation tube 101 along with other cell fluids, namely, the separation of target cells and the feedback of blood are realized. After the separated cellular fluid is output from the separation tube 101, the annular magnetic separation unit 105 is separated from the second surface 1042, as shown in fig. 5, for example, the separation tube 101 is inverted to make the second surface 1042 below the first surface 1041, and under the action of gravity, the annular magnetic separation unit 105 is separated from the second surface 1042 of the piston 104, at this time, the magnetic beads combined with the CD34 cells are no longer adsorbed by the magnetic force and fall off from the first surface 1041, and the CD34 cells can be left in the separation tube 101.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.

Claims

1. A cell separation and reinfusion device, comprising: the cell separation device comprises a cylindrical separation tube, wherein a movable assembly in sliding connection with the separation tube is sleeved in the separation tube, the movable assembly comprises a piston shaft, one end of the piston shaft is fixedly connected with a piston, the piston comprises a first surface and a second surface which are parallel to each other, the first surface and the second surface are perpendicular to the axial direction of the piston shaft, a cavity for containing cell liquid to be separated is formed between the first surface and the inner surface of the separation tube, the cell liquid to be separated comprises target cells, an annular magnetic separation unit in sliding connection with the piston shaft is further sleeved on the piston shaft, the annular magnetic separation unit is positioned on one side close to the second surface of the piston and is parallel to the second surface of the piston, a first pipeline and a second pipeline which are communicated with the cavity are further arranged on the separation tube, and the first pipeline is used for inputting the cell liquid to be separated, the second pipeline is used for inputting magnetic substances combined with target cells, and the annular magnetic separation unit is in contact connection with the second surface and adsorbs the magnetic substances combined with the target cells onto the first surface by means of magnetic force.

2. The cell separation and feedback device according to claim 1, wherein: one end of the separation tube is provided with an opening, the other end of the separation tube is provided with a blocking piece for blocking, and the piston shaft and the piston sleeve are arranged in the opening.

3. The cell separation and feedback device according to claim 2, wherein: the separation blade is provided with a first through hole and a second through hole, and the first pipeline and the second pipeline are respectively communicated with the first through hole and the second through hole.

4. The cell separation and feedback device according to claim 1, wherein: one end of the first pipeline, which is far away from the separation pipe, is connected with a collection device and/or a liquid outlet pipe for collecting cell sap to be separated.

5. The cell separation and feedback device according to claim 1, wherein: one end of the second pipeline, which is far away from the separation pipe, is connected with a liquid inlet pipe and/or a bottle stopper puncture outfit.

6. The cell separation and feedback device according to claim 1, wherein: and the first pipeline and/or the second pipeline are/is provided with a control clamp for controlling the flow.

7. The cell separation and feedback device according to claim 1, wherein: one end of the first pipeline and/or the second pipeline, which is far away from the separation pipe, is connected with a luer.

8. The cell separation and feedback device according to claim 1, wherein: the magnetic substance is an antibody-coated magnetic bead.

9. The cell separation and feedback device according to claim 1, wherein: the first pipeline and the second pipeline are parallel to the axial direction of the separation pipe.

10. The cell separation and feedback device according to claim 1, wherein: and one end of the piston shaft, which is far away from the piston, is fixedly connected with a piston handle.