Trap type centrifugal tube for primary cell separation

Abstract

The invention relates to the technical field of centrifugal tubes, in particular to a trap type centrifugal tube for primary cell separation. According to the technical scheme, the centrifugal tube comprises a centrifugal tube main body, wherein the centrifugal tube main body is sequentially provided with a thick tube part, a conical part and a thin tube part from top to bottom; the thin pipe part is provided with a plurality of prefabricated fractures in an array mode in the height direction of the thin pipe part; the conical part is used for communicating the thick pipe part with the thin pipe part, and the section of the conical part is funnel-shaped; a thin tube bottom with a semi-arc-shaped section is arranged at the bottom of the thin tube part; supporting side wings arranged on the outer wall of the thin pipe part in an array mode are arranged between the prefabricated fractures of each layer. And the sizes of the supporting side wings at the lower end of the thin tube part are gradually reduced. The method is relatively simple and convenient to operate, requires little time, does not need additional solution or antigen, and improves and avoids the influence of non-culture medium solution on cells.

Description

Technical Field

[0001] The invention relates to the technical field of centrifuge tubes, in particular to a trap-type centrifuge tube for separating primary cells. Background Art

[0002] When primary cells are separated, they are often mixed with various cell groups in the tissue. There are various separation methods to provide different levels and purification solutions for cell separation according to different research and application requirements. Perco ll density gradient centrifugation is mainly based on the suspension of cells to separate the required cells. Using this method, it is necessary to prepare a Perco ll separation solution of corresponding concentration, and it is necessary to prepare two or more Perco ll separation solutions of different concentrations, so that the required cells are between two Perco ll separation solutions after centrifugation, and then the middle layer cells are aspirated for subsequent culture and other purposes. The principle adopted by immunomagnetic bead cell sorting technology is mainly to combine the antigens on the cell surface with the specific antibodies carried by the magnetic beads. Immunomagnetic beads are divided into positive selection and negative selection. Among them, Perco ll is mainly used for preliminary rough separation based on cell density (suspension), while methods such as immunomagnetic beads mainly use antibodies to finely separate cells containing specific surface proteins.

[0003] Perco ll separation solution requires a specific solvent when preparing, and the use and preparation of different brands of Perco ll cannot be unified. The density range of Perco ll separation solution is limited. If the suspension density of the cell is greater than the maximum limit or lower than the minimum limit, it cannot be separated by this method. When using the Perco ll separation method, other cells will exist in the middle layer, and the required cells cannot be accurately separated. And Perco ll itself is a colloidal liquid. After separating the required cells, it needs to be washed with culture medium again, and the required cells will be lost to a certain extent. The immunomagnetic bead sorting technology requires the use of magnetic beads and magnets that are compatible with the cells, and it is necessary to purchase antibodies corresponding to the cell surface. The separation technology and cost are relatively high, and this method is not suitable for batch separation of primary cells. In addition, the immunomagnetic bead separation method is extremely expensive, and the load capacity for the separated cells is extremely limited. It usually requires a rough separation in advance to increase the separation purity and separation efficiency of such methods as magnetic bead separation. In view of the above reasons, the present application proposes a trap-type centrifuge tube for primary cell separation that does not require the use of additional solutions or antigen antibodies. Summary of the invention

[0004] The purpose of the present invention is to address the problems existing in the background technology and to provide a trap-type centrifuge tube for primary cell separation that does not require the aid of additional solutions or antigens and antibodies.

[0005] The technical solution of the present invention is: a trap-type centrifuge tube for primary cell separation, comprising a centrifuge tube body, wherein the centrifuge tube body is provided with a thick tube portion, a tapered portion and a thin tube portion in sequence from top to bottom;

[0006] Wherein, the thin tube part is provided with a plurality of prefabricated fractures in an array along its height direction.

[0007] Optionally, the tapered portion is used to connect the thick tube portion and the thin tube portion, and the cross-section of the tapered portion is funnel-shaped.

[0008] Optionally, the bottom of the capillary portion is provided with a capillary bottom having a semicircular arc cross-section.

[0009] Optionally, supporting side wings arranged in an array on the outer wall of the capillary portion are provided between the prefabricated fractures of each layer.

[0010] Optionally, the lateral width of the supporting wing is arranged in a decreasing manner from top to bottom along the length direction of the thin tube portion, and forms an inverted triangle structure at the bottom of the thin tube portion.

[0011] Optionally, the centrifuge tube body is snap-fitted and adapted to the inner wall of a 15 ml external centrifuge tube, and the centrifuge tube body is sleeved inside the external centrifuge tube.

[0012] Compared with the prior art, the present invention has the following beneficial technical effects:

[0013] The present invention does not require the use of additional solutions or antigens and antibodies. It only requires adding a cell suspension of a certain cell concentration into the trap-type centrifuge tube when separating cells to separate specific cells. The operation is relatively simple, time-consuming, and the efficiency of obtaining crudely separated cell groups is high and the cost is low.

[0014] Furthermore, the present invention is adapted to intercepting capillaries of different heights for separation by setting the prefabricated fracture, and each section of the cell suspension in the capillaries is placed in a dish for culture;

[0015] Furthermore, the separation methods used in the present invention are all physical separation, which does not affect the physical and chemical properties of cells, can maintain the characteristics of primary cells, and can separate primary cells of most tissue types. If a large number of primary cells need to be separated, several preliminary experiments can be performed to determine the position of the required cells in the capillary tube, and then the primary cells can be separated in batches;

[0016] In summary, the present invention is relatively simple to operate, requires very little time, does not require additional solutions or antigens, and improves the avoidance of non-culture medium solutions from influencing cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 Provide a front view schematic diagram of the present invention;

[0018] Figure 2 for Figure 1 Schematic cross-section of the middle LL;

[0019] Figure 3 for Figure 1 A schematic diagram of the main view;

[0020] Figure 4 for Figure 1 Schematic diagram of the cross section at AA in the middle;

[0021] Figure 5 for Figure 1 Schematic diagram of the cross section at CC in the middle;

[0022] Figure 6 for Figure 1 Schematic diagram of the cross section at EE in the middle;

[0023] Figure 7 for Figure 1 Schematic diagram of the cross-section front view at FF in the middle.

[0024] Reference numerals:

[0025] 1. Centrifuge tube body; 11. Thick tube part; 12. Conical part; 13. Prefabricated fracture; 14. Support wing; 15. Thin tube part; 16. Thin tube bottom. DETAILED DESCRIPTION

[0026] The technical solution of the present disclosure will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present disclosure, rather than all of the embodiments.

[0027] The components of the embodiments of the present disclosure generally described and shown in the drawings herein may be arranged and designed in a variety of different configurations. Therefore, the following detailed description of the embodiments of the present disclosure provided in the drawings is not intended to limit the scope of the present disclosure claimed, but merely represents selected embodiments of the present disclosure.

[0028] Based on the embodiments in the present disclosure, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present disclosure.

[0029] In the description of the present disclosure, it should be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present disclosure and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present disclosure.

[0030] In the description of the present disclosure, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present disclosure can be understood according to specific circumstances.

[0031] like Figure 1-Figure 7 As shown, the present invention proposes a trap-type centrifuge tube for primary cell separation, including a centrifuge tube body 1, which is clamped and adapted to the inner wall of a 15ml external centrifuge tube, and the centrifuge tube body 1 is sleeved inside the external centrifuge tube, and then placed in a centrifuge for centrifugation. Specifically, the centrifuge tube body 1 is provided with a thick tube portion 11, a tapered portion 12 and a thin tube portion 15 from top to bottom, and the tapered portion 12 is used to connect the thick tube portion 11 and the thin tube portion 15; and the cross section of the tapered portion 12 is funnel-shaped.

[0032] Among them, the capillary portion 15 is arrayed with a plurality of prefabricated fractures 13 along its height direction, and between each layer of prefabricated fractures 13 there are support side wings 14 arranged in an array on the outer wall of the capillary portion 15, and the lateral width of the support side wings 14 is arranged in a decreasing manner from top to bottom along the length direction of the capillary portion 15, and is located at the bottom of the capillary portion 15 to form an inverted triangle structure; the bottom of the capillary portion 15 is provided with a capillary bottom 16 with a semicircular arc cross-section.

[0033] In this embodiment, the cell separation operation is carried out according to the following steps:

[0034] 1. Add a certain amount of culture medium to the trap centrifuge tube, fill the capillary portion 15 and the upper portion, remove all bubbles in the capillary tube, and add the same culture medium at the conical portion 12 at the upper portion of the centrifuge tube;

[0035] 2. Take 500 μL of completely mixed cell suspension as a centrifugal portion as an example, slightly tilt the trap-type centrifuge tube of the present application, and slowly add the cell suspension to the upper layer of the culture medium along the tube wall, trying not to affect the stratification of the lower layer. Each time the cell suspension is added, the cell suspension in the original centrifuge tube must be blown and mixed;

[0036] 3. Put the trap centrifuge tube into a 15mL centrifuge tube and put it into the centrifuge for centrifugation;

[0037] 4. After centrifugation, aspirate the liquid above the capillary portion 15;

[0038] 5. Then, take out the trap centrifuge tube from the 15 mL centrifuge tube, and then use tweezers and scissors to cut the thin tube containing cells from top to bottom at the pre-made fracture position 13;

[0039] 6. Place the capillaries containing cells in different culture wells, and use culture medium to wash the cells in the capillaries into culture dishes for culture;

[0040] 7. Repeat the above two steps until every section of the cell suspension in the capillary tube is placed in a dish for culture.

[0041] It should be noted in the present application that the cells in the separated tissue should be digested into single cells as much as possible and should not be aggregated into clumps, otherwise the cell clumps will block the mouth of the capillary tube during centrifugation, making it impossible for the cells to enter the prefabricated fracture 13; the amount of cells required for a trap-type centrifuge tube must reach or exceed the total volume of the capillary portion 15 to meet the conditions of use; when adding the cell suspension to the trap-type centrifuge tube, it should be added slowly to the upper layer of the culture medium, trying not to affect the lower separation layer; after centrifugation, the cells should fill the prefabricated fracture 13, or only exist in the prefabricated fracture 13, so as to achieve a better separation effect; it should be noted that the break of each layer of the prefabricated fracture 13 needs to be cut by an adapted pliers, and the pliers are provided with an opening in the shape of a corresponding supporting wing 14, and the corresponding prefabricated fracture 13 can be clamped after the corresponding supporting wing 14 is stuck; when harvesting cells, the culture medium in the thick tube portion 11 is first sucked out, and then the capillary portions 15 of different heights are cut off respectively.

[0042] The above specific embodiments are merely optional embodiments of the present invention. Based on the technical solutions of the present invention and the relevant inspirations of the above embodiments, those skilled in the art may make various alternative improvements and combinations to the above specific embodiments.

Claims

1. A trap-type centrifuge tube for primary cell separation, characterized in that: The centrifuge tube body (1) comprises a thick tube portion (11), a tapered portion (12) and a thin tube portion (15) arranged in sequence from top to bottom; Wherein, the thin tube portion (15) is provided with a plurality of prefabricated fractures (13) in an array along its height direction.

2. A trap-type centrifuge tube for primary cell separation according to claim 1, characterized in that: The tapered portion (12) is used to connect the thick tube portion (11) and the thin tube portion (15), and the cross section of the tapered portion (12) is a funnel-shaped structure.

3. A trap-type centrifuge tube for primary cell separation according to claim 1, characterized in that: The bottom of the thin tube portion (15) is provided with a thin tube bottom (16) having a semicircular arc cross section.

4. A trap-type centrifuge tube for primary cell separation according to claim 1, characterized in that: Support side wings (14) arranged in an array on the outer wall of the thin tube portion (15) are provided between the prefabricated fractures (13) of each layer.

5. A trap-type centrifuge tube for primary cell separation according to claim 4, characterized in that: The lateral width of the supporting side wings (14) is arranged in a decreasing manner from top to bottom along the length direction of the thin tube portion (15), and is located at the bottom of the thin tube portion (15) to form an inverted triangle structure.

6. A trap-type centrifuge tube for primary cell separation according to claim 1, characterized in that: The centrifuge tube body (1) is fitted and matched with the inner wall of an external centrifuge tube, and the centrifuge tube body (1) is sleeved inside the external centrifuge tube.

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