Coated leukocyte single liquid collection container for circulating tumor cell sorting

Abstract

A coating leukocyte single collection liquid container for circulating tumor cell sorting comprises a shell, a mixing cavity and a cell collection cavity are arranged in the shell, the mixing cavity is provided with a cell sample input port and a cryoprotectant input port, the cell collection cavity is arranged on the lower side of the mixing cavity, and the cryoprotectant input port is arranged on the lower side of the cell collection cavity. The inner wall of the mixing cavity and the inner wall of the cell collecting cavity are sequentially coated with a base layer and a functional layer from inside to outside, the base layer is made of polyethylene glycol, the functional layer is made of modified polyvinylpyrrolidone, and the shell is made of polytetrafluoroethylene. By applying the modified polyvinylpyrrolidone, surface negative charges can be effectively neutralized, non-specific adsorption of the CTCs is effectively reduced, the original physiological state of the CTCs is kept, the efficiency and accuracy of CTCs sorting are improved, and the clinical requirements of precise medical treatment and liquid biopsy are met; the mixing cavity and the cell collecting cavity are arranged in the container, the leukocyte single collection liquid and the cryoprotectant are fully mixed in the mixing cavity and then enter the cell collecting cavity, then the whole container is cryopreserved, and operation is convenient.

Description

Technical Field

[0001] The present invention relates to the field of chemistry, particularly to experimental equipment, and more particularly to a coated leukapheresis fluid container for circulating tumor cell sorting. Background Art

[0002] Circulating tumor cells (CTCs) are tumor cells that shed from primary or metastatic tumors and enter the peripheral blood circulation, and are important biomarkers for cancer diagnosis and treatment. Leukapheresis is a technique for separating and collecting mononuclear cells (including white blood cells and CTCs) from whole blood by a blood component separator. A leukapheresis fluid container is a special container for collecting and storing leukapheresis products.

[0003] Currently, the main challenge in CTC detection is that their quantity in peripheral blood is extremely small, and the detection rate of traditional small-volume blood sample (7.5 - 10 mL) detection methods is low. Research shows that processing a large amount of blood through leukapheresis technology can significantly improve the detection rate of CTCs, and the detection rate can reach over 90% in non-metastatic breast cancer patients.

[0004] However, the surface of the existing leukapheresis fluid container has a negative charge property, which can cause non-specific adsorption of CTCs, affecting their physiological state and subsequent analysis; existing container materials such as PVC are prone to becoming brittle at low temperatures and are not suitable for cryopreservation of CTCs; the surface of existing container materials promotes cell adhesion, which is not conducive to the complete recovery of CTCs. When storing leukapheresis fluid, it is necessary to first mix it with a cryoprotectant and then take out the mixed liquid for cryopreservation, which is inconvenient to operate. Summary of the Invention

[0005] The purpose of the present invention is to provide a coated leukapheresis fluid container for circulating tumor cell sorting, and the such coated leukapheresis fluid container for circulating tumor cell sorting of the present invention aims to solve the above technical problems existing in the existing leukapheresis fluid container.

[0006] The such coated leukapheresis fluid container for circulating tumor cell sorting of the present invention includes a housing. A mixing chamber and a cell collection chamber are arranged inside the housing. The mixing chamber is provided with a cell sample input port and a cryoprotectant input port. The cell collection chamber is arranged below the mixing chamber. A channel is arranged between the mixing chamber and the cell collection chamber. A groove horizontally passing through the channel is opened on the housing, and a plug board is inserted into the groove. The inner walls of the mixing chamber and the cell collection chamber are coated with a base layer and a functional layer in sequence from the inside to the outside. The material of the base layer is polyethylene glycol, the material of the functional layer is modified polyvinylpyrrolidone, and the material of the housing is polytetrafluoroethylene.

[0007] Further, the surface of the functional layer is coated with an anti-adhesion layer, and the material of the anti-adhesion layer is phosphatidylcholine polymer.

[0008] Further, a partition is provided in the housing between the mixing chamber and the cell collection chamber. The channel is vertically provided in the partition. The channel communicates the mixing chamber and the cell collection chamber, and the groove extends into the partition.

[0009] Further, the upper side surface of the partition is provided with a V-shaped groove, and the channel is located at the bottom of the V-shaped groove.

[0010] Further, a draw groove is provided in the cell collection chamber. The material of the draw groove is polytetrafluoroethylene. The draw groove is provided with a storage chamber with an upward opening. The surface of the storage chamber is coated with a base layer and a functional layer in sequence from the inside to the outside. The surface of the functional layer is coated with an anti-adhesion layer, and the material of the anti-adhesion layer is phosphatidylcholine polymer.

[0011] Further, a groove is provided in the housing around the top of the cell collection chamber. A sealing ring is provided in the groove. The lower end of the sealing ring is hermetically connected to the upper end of the draw groove. A handle is provided on one side of the draw groove.

[0012] Further, the number of the storage chambers is multiple, the number of the channels is the same as the number of the storage chambers, and the lower end of the channel corresponds to the opening position of the storage chamber.

[0013] Compared with the prior art, the effect of the present invention is positive and obvious. The present invention applies modified polyvinylpyrrolidone as a hydrophilic coating on the inner surface of the traditional leukapheresis liquid container, which can effectively neutralize the surface negative charge, effectively reduce the non-specific adsorption of CTCs, maintain the original physiological state of CTCs, improve the efficiency and accuracy of CTC sorting, and meet the clinical needs of precision medicine and liquid biopsy; a mixing chamber and a cell collection chamber are provided inside the container. After the leukapheresis liquid and the cryoprotectant are fully mixed in the mixing chamber, the fluid on-off mechanism is opened and flows into the cell collection chamber, and then the whole container is frozen and stored, which is convenient to operate. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 is a schematic structural diagram of the present invention.

[0015] Figure 2 is a schematic internal structural diagram of the present invention.

[0016] Figure 3 is a schematic internal side view structural diagram of the present invention.

[0017] Figure 4 is Figure 2 a cross-sectional view of the coating structure at A in

[0018] Figure 5Yes Figure 2 Cross-sectional view of the coating structure at position B in

[0019] Markings in the figure: 1. Shell; 2. Cell collection chamber; 3. Mixing chamber; 4. Cryoprotectant inlet; 5. Cell sample inlet; 6. Channel; 7. Groove; 8. Insertion plate; 9. Pulling slot; 10. Sealing ring; 11. Storage chamber; 12. Handle; 13. Base layer; 14. Functional layer; 15. Anti-adhesion layer. Detailed implementation mode

[0020] The present invention will be further described below in conjunction with the drawings and embodiments, but the present invention is not limited thereby.

[0021] Embodiment 1

[0022] As Figures 1-4 shown, a coating leukapheresis liquid container for circulating tumor cell sorting of the present invention includes a shell 1, a mixing chamber 3 and a cell collection chamber 2 are arranged in the shell 1, a cell sample inlet 5 and a cryoprotectant inlet 4 are arranged in the mixing chamber 3, the cell collection chamber 2 is arranged below the mixing chamber 3, a channel 6 is arranged between the mixing chamber 3 and the cell collection chamber 2, a groove 7 horizontally passing through the channel 6 is opened on the shell 1, an insertion plate 8 is inserted in the groove 7, the inner walls of the mixing chamber 3 and the cell collection chamber 2 are coated with a base layer 13 and a functional layer 14 in sequence from inside to outside, the material of the base layer 13 is polyethylene glycol, the material of the functional layer 14 is modified polyvinylpyrrolidone, and the material of the shell 1 is polytetrafluoroethylene.

[0023] Furthermore, a partition is arranged between the mixing chamber 3 and the cell collection chamber 2 of the shell 1, the channel 6 is vertically arranged in the partition, the channel 6 communicates the mixing chamber 3 and the cell collection chamber 2, and the groove 7 extends into the partition.

[0024] Furthermore, the upper side of the partition is set as a V-shaped groove, and the channel 6 is located at the bottom of the V-shaped groove.

[0025] Specifically, the mixing chamber 3 is used for mixing the cell sample and the cryoprotectant. When the insertion plate 8 is inserted into the groove 7, the channel 6 is blocked. After the cell sample and the cryoprotectant are mixed, the insertion plate 8 is pulled out. One end of the insertion plate 8 extends out of the shell 1 for convenient operation from the outside. The mixed liquid is input into the cell collection chamber 2 through the channel 6 for storage, and then the insertion plate 8 is inserted into the groove 7 to seal the cell collection chamber 8 from the outside. The base layer 13 is made of polyethylene glycol material, which can effectively improve the compatibility between the inner cavity and the cell sample. The functional layer is made of modified polyvinylpyrrolidone material, which has excellent hydrophilicity and the characteristic of reducing cell adhesion, can effectively reduce the non-specific adsorption of CTCs, improve the detection rate of CTCs, and the shell 1 is made of polytetrafluoroethylene material, which has excellent low-temperature stability.

[0026] Furthermore, a draw slot 9 is provided in the cell collection chamber 2. The draw slot 9 is made of polytetrafluoroethylene. The draw slot 9 is provided with a storage chamber 11 with an upward opening. The surface of the storage chamber 11 is coated with a base layer 13 and a functional layer 14 in sequence from the inside to the outside.

[0027] Specifically, when setting the draw slot 9 to facilitate subsequent extraction of cell samples for analysis, detection or clinical applications, the draw slot 9 can be directly pulled out to take out the sample stored in the storage chamber 11. As Figure 5 shown, coating the base layer 13, the functional layer 14 and the anti-adhesion layer 15 on the surface of the draw slot 9 has the same effect as the base layer 13 and the functional layer 14 of the housing 1.

[0028] Furthermore, a groove 7 is provided in the housing 1 around the top of the cell collection chamber 2. A sealing ring 10 is provided in the groove 7. The lower end of the sealing ring 10 is sealingly connected to the upper end of the draw slot 9. A handle 12 is provided on one side of the draw slot 9.

[0029] Specifically, when the draw slot 9 is inside the cell collection chamber 2, the sealing ring 10 in the groove 7 seals the top of the draw slot 9, isolating the storage chamber 11 from the outside and preventing the leukapheresis liquid in the storage chamber 11 from overflowing. The handle 12 is provided to facilitate pulling out the draw slot 9.

[0030] Furthermore, the number of the storage chambers 11 is multiple, and the number of the channels 6 is the same as that of the storage chambers 11. The lower end of the channel 6 corresponds to the opening position of the storage chamber 11.

[0031] Specifically, setting multiple storage chambers 11 can separately store multiple samples for backup.

[0032] When this embodiment is in use, first inject the leukapheresis liquid or blood sample into the mixing chamber 3 of the housing 1 through the cell sample input port 5, and add an appropriate amount of cryoprotectant through the cryoprotectant 4 input port as needed, and mix well with the sample in the mixing chamber 3. Then pull the insertion plate 8 to open the channel 6. The mixed leukapheresis liquid enters the storage chamber 11 of the draw slot 9 through the channel 6, and then push the insertion plate 8 to close the channel 6. The leukapheresis liquid is stored in the storage chamber 11 of the draw slot 9. The sealing ring 10 prevents the leukapheresis liquid from overflowing, facilitating subsequent cryopreservation or transportation. When analysis and detection are needed, pull the handle 12 to pull out the draw slot 9 to take out the sample.

[0033] The housing 1 is made of polytetrafluoroethylene (PTFE) material, enabling the container to maintain good flexibility and stability under ultra-low temperature conditions of -196 °C, and it will not become brittle like traditional PVC materials. The base layer 13 is a polyethylene glycol (PEG) base layer, which can effectively neutralize the surface negative charges, reduce the non-specific adsorption of CTCs, and maintain the original morphology and function of CTCs. The functional layer 14 is made of modified polyvinylpyrrolidone (PVP), which provides a "slippery" surface, reducing the friction coefficient between CTCs and the container surface by 85%, and greatly improving the recovery rate of CTCs. The CAS number of the modified polyvinylpyrrolidone is 9003-39-8, the molecular formula is (C6H9NO)n, and the structural formula is:

[0034] Purchased from BASF Corporation in Germany (PVPK30K--90).

[0035] Example 2

[0036] On the basis of Example 1, an anti-adhesion layer 15 is coated on the surface of the functional layer 14, and the material of the anti-adhesion layer 15 is phosphatidylcholine polymer.

[0037] Specifically, the CAS number of the phosphatidylcholine polymer monomer is 16904-96-4,

[0038] The structural formula is:

[0039] (Purchased from the agent: Shanghai Dongtu Chemical Industry Import and Export Co., Ltd.); The phosphatidylcholine polymer has excellent anti-pollution performance, can effectively inhibit microbial adhesion and biofilm formation, reducing the microbial adhesion rate by 95%, significantly reducing the pollution risk of the sample; improving the biocompatibility of the inner surface of the container by 40%, making it closer to the natural environment in the human body and reducing the stimulation to CTCs.

Claims

1. A coated leukapheresis fluid container for circulating tumor cell sorting, characterized in that: It includes a housing (1), a mixing chamber (3) and a cell collection chamber (2) are arranged inside the housing (1), the mixing chamber (3) is provided with a cell sample inlet (5) and a cryoprotectant inlet (4), the cell collection chamber (2) is arranged on the lower side of the mixing chamber (3), a channel (6) is arranged between the mixing chamber (3) and the cell collection chamber (2), the housing (1) is provided with a groove (7) that horizontally penetrates the channel (6), and a plug board (8) is inserted in the groove (7). The inner walls of the mixing chamber (3) and the cell collection chamber (2) are coated with a base layer (13) and a functional layer (14) in sequence from the inside to the outside. The material of the base layer (13) is polyethylene glycol, the material of the functional layer (14) is modified polyvinylpyrrolidone, and the material of the housing (1) is polytetrafluoroethylene.

2. The coated leukapheresis fluid container for circulating tumor cell sorting according to claim 1, wherein: The surface of the functional layer (14) is coated with an anti-adhesion layer (15), and the material of the anti-adhesion layer (15) is phosphatidylcholine polymer.

3. The coated leukapheresis fluid container for circulating tumor cell sorting according to claim 1, wherein: A pull-out groove (9) is arranged inside the cell collection chamber (2), the material of the pull-out groove (9) is polytetrafluoroethylene, the pull-out groove (9) is provided with a storage chamber (11) with an upward opening, and the surface of the storage chamber (11) is coated with a base layer (13) and a functional layer (14) in sequence from the inside to the outside. The surface of the functional layer (14) is coated with an anti-adhesion layer (15), and the material of the anti-adhesion layer (15) is phosphatidylcholine polymer.

4. The coated leukapheresis fluid container for circulating tumor cell sorting according to claim 3, characterized in that: A groove (7) is arranged in the housing (1) around the top of the cell collection chamber (2), a sealing ring (10) is arranged in the groove (7), the lower end of the sealing ring (10) is hermetically connected to the upper end of the pull-out groove (9), and a handle (12) is arranged on one side of the pull-out groove (9).

5. The coated leukapheresis fluid container for circulating tumor cell sorting according to claim 3, characterized in that: The number of the storage chambers (11) is multiple, the number of the channels (6) is the same as the number of the storage chambers (11), and the lower end of the channel (6) corresponds to the opening position of the storage chamber (11).

6. The coated leukapheresis fluid container for circulating tumor cell sorting according to claim 1, characterized in that: A partition is arranged between the mixing chamber (3) and the cell collection chamber (2) in the housing (1), the channel (6) is vertically arranged in the partition, the channel (6) communicates the mixing chamber (3) and the cell collection chamber (2), and the groove (7) extends into the partition.

7. The coated leukapheresis fluid container for circulating tumor cell sorting according to claim 6, wherein: The upper side of the partition is arranged as a V-shaped groove, and the channel (6) is located at the bottom of the V-shaped groove.

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