CN115968865A

CN115968865A - CAR-T cell cryopreservation box and cryopreservation method

Info

Publication number: CN115968865A
Application number: CN202310048691.1A
Authority: CN
Inventors: 韩刚; 王全军; 熊克朝; 刘杨
Original assignee: Beijing Saifu Pharmaceutical Research Institute Co ltd
Current assignee: Beijing Saifu Pharmaceutical Research Institute Co ltd
Priority date: 2023-02-01
Filing date: 2023-02-01
Publication date: 2023-04-18
Anticipated expiration: 2043-02-01
Also published as: CN115968865B

Abstract

The invention belongs to the technical field of cryopreservation boxes, in particular to a CAR-T cell cryopreservation box and a cryopreservation method, aiming at the problems that in the prior art, cold air in the cryopreservation box is easy to overflow and scatter, and when a plurality of cryopreservation tubes need to be placed, the cryopreservation tubes placed in an early stage gradually lose the freezing effect, the invention provides the following scheme, which comprises the following steps: the freezing storage box is hinged to the top of the freezing storage box, hinge pins are fixedly connected to two sides of the freezing storage box, one end, far away from each other, of each hinge pin is rotatably connected with the same handle, a sliding plate is connected in the freezing storage box in a sliding mode, in the freezing storage box, the sealing of the sealing plate on the first through hole can be removed through the dead weight of the freezing storage tube and CAR-T cell suspension liquid, so that the freezing storage tube is preliminarily frozen, the sliding plate moves downwards for a small distance along with the increase of the number of the freezing storage tubes, air holes are opened, freezing gas in the liquid nitrogen bin freezes the freezing storage tube, and the freezing storage tube placed in the early stage can be continuously frozen without overflowing of the freezing gas in the liquid nitrogen bin.

Description

CAR-T cell cryopreservation box and cryopreservation method

Technical Field

The invention relates to the technical field of cryopreservation boxes, in particular to a CAR-T cell cryopreservation box and a cryopreservation method.

Background

The CAR-T cell has the characteristics completely different from other non-in-vivo amplification medicaments, and is characterized in that individuation, small yield, limited batch and the like are reflected, so that a personal seed bank is established in a targeted manner to provide a stable source for long-term production, and the CAR-T cell storage is performed by using a CAR-T special cryopreservation box.

The cryopreservation box in the prior art still has the following problems when in use:

1. in the prior art, when CAR-T cells are extracted, the CAR-T cells are required to be prepared into suspension liquid and then are injected into a cryopreservation tube and are placed in a cryopreservation box for storage, but when the cryopreservation box is opened, cold air in the cryopreservation box overflows in the process of placing the cryopreservation tube, so that the later cryopreservation effect is influenced;

2. when a large number of cryopreservation tubes need to be placed, the early-stage placed cryopreservation tubes gradually lose the freezing effect, and the CAR-T cell suspension in the cryopreservation tubes is easy to deteriorate.

In order to solve the problems, the invention provides a CAR-T cell cryopreservation box and a cryopreservation method.

Disclosure of Invention

The invention provides a CAR-T cell cryopreservation box and a cryopreservation method, and overcomes the defects that in the prior art, cold air in the cryopreservation box is easy to overflow and scatter, and when a large number of cryopreservation tubes need to be placed, the cryopreservation tubes placed in the early stage gradually lose the freezing effect.

The invention provides the following technical scheme:

a CAR-T cell cryopreservation cassette comprising: the top of the freezing storage box is hinged with a cover plate, two sides of the freezing storage box are fixedly connected with pin shafts, and one ends of the two pin shafts, which are far away from each other, are rotatably connected with the same handle;

the freezing storage box is internally and fixedly connected with a fixed plate positioned below the sliding plate in a sliding manner, and the sliding plate is elastically connected with the fixed plate through a buffer structure;

the pre-freezing structure is arranged in the freezing storage box and is used for primarily freezing the freezing storage pipe;

the gas structure is frozen in the expansion, sets up in the fixed plate for the volume of gas is frozen in the increase, is convenient for freeze the pipe fast and freezes the deposit.

In one possible design, the pre-freezing storage structure comprises a top plate fixedly connected in a freezing storage box, a plurality of freezing storage sleeves are fixedly arranged in the top plate in a penetrating mode, sliding rods are arranged on the inner walls of the bottoms of the freezing storage sleeves in a sliding penetrating mode, trays located in the freezing storage sleeves are fixedly connected to the top ends of the sliding rods, transverse plates are fixedly connected to the bottom ends of the sliding rods, sealing plates are fixedly connected to the two sides of the top of each transverse plate, a plurality of first through holes are arranged on the two sides of each freezing storage sleeve, a plurality of second through holes matched with the first through holes are arranged in the sealing plates, the first through holes and the second through holes are mutually staggered under normal conditions, the first through holes can be sealed through the sealing plates, frozen gas in the pre-freezing storage cavities can be prevented from entering the freezing storage sleeves to cause overflowing of the frozen gas, when the freezing storage tubes are placed in the freezing storage sleeves, the sliding rods and the sealing plates are pushed by the gravity of the freezing storage tube suspension liquid nitrogen-storing cell and the liquid nitrogen, the first through holes are aligned with the second through holes at the moment, and then the frozen gas diffused in the freezing storage box is absorbed into the pre-freezing storage box to initially to carry out the liquid nitrogen storage sliding plate.

In a possible design, the structure for expanding frozen gas comprises a plurality of vent holes arranged in a fixed plate, the top of the fixed plate is connected with a sliding closed block used for closing the vent holes in a sliding mode, the top of the sliding closed block is connected with a connecting rod in a rotating mode, the top end of the connecting rod is connected with the bottom of a sliding plate in a rotating mode, when the number of the frozen pipes placed in a freezing storage sleeve is increased, the transverse plates are driven by the gravity of the plurality of frozen pipes to downwards extrude the sliding plate, the sliding plate downwards moves for a small distance, the sliding plate drives the connecting rod to rotate, and the connecting rod pushes the sliding closed block to move for a certain distance, so that the blocking of the sliding closed block on the vent holes is gradually removed, frozen gas in a liquid nitrogen bin enters a pre-freezing storage cavity through the vent holes and round holes in the sliding plate, the frozen gas in the pre-freezing storage cavity is increased, the frozen pipes placed in the liquid nitrogen bin are further frozen, namely, the frozen gas in the liquid nitrogen bin is prevented from overflowing quickly, the frozen pipes in the liquid nitrogen bin can be frozen pipes, and the frozen pipes can be frozen before, and the frozen pipes are prevented from going bad.

In a possible design, the top of the sliding plate is fixedly connected with a plurality of first heat insulation cylinders, the inner wall of the bottom of each first heat insulation cylinder is fixedly connected with a first spring, each first heat insulation cylinder is slidably connected with a first fixing column fixedly connected with the top end of the corresponding first spring, the top end of each first fixing column is fixedly connected with the bottom of the corresponding transverse plate, and through the matching of the corresponding first fixing column and the corresponding first fixing column, the sealing of the corresponding first through hole by the corresponding sealing plate can be removed according to the gravity of the cryopreserved pipe and the CAR-T cell suspension liquid, so that the phenomenon that frozen gas overflows and scatters through the corresponding first through hole can be avoided, and the cryopreserved pipe can be frozen in time.

In a possible design, the buffer structure comprises a plurality of second temperature-isolating cylinders fixedly connected to the top of the fixing plate, second springs are fixedly connected to the inner walls of the bottoms of the second temperature-isolating cylinders, the elastic force of the second springs is larger than that of the first springs, second fixing columns fixedly connected to the top ends of the second springs are slidably connected to the second temperature-isolating cylinders, the top ends of the second fixing columns are fixedly connected to the bottom of the sliding plate, the sliding plate can be prevented from moving downwards when the number of the cryopreservation tubes is small through the cooperation of the second springs and the second fixing columns, the vent holes are prevented from being opened, when the number of the cryopreservation tubes is small, the sliding plate can push the second fixing columns to move downwards, the vent holes are gradually opened, and then more frozen gas can enter the pre-cryopreservation cavity to freeze the cryopreservation tubes.

In a possible design, the fixed plate forms the liquid nitrogen storehouse with the bottom inner wall that freezes deposits the box, the top of sliding plate and the bottom of roof form in advance to freeze and deposit the chamber, and freeze and deposit telescopic bottom and extend to in freezing in advance and deposit the chamber, be equipped with a plurality of round holes in the sliding plate, freezing gas in the liquid nitrogen storehouse permeates the back penetration after the heat preservation of sliding plate and fixed plate and freezes in advance and deposit the chamber, just in freezing in advance and deposit remaining freezing gas in the chamber can tentatively freeze the pipe and deposit to freezing, can avoid the excessive scattering of freezing gas in the liquid nitrogen storehouse again.

In a possible design, it has a plurality of catch bars to slide to run through in the roof, and the bottom of catch bar and the top fixed connection of sliding plate, be equipped with U type hole in the catch bar, and two upper and lower through-holes in U type hole are located the upper and lower position of roof respectively, promote the catch bar at the apron and move the back down, the hole at the both ends in U type hole just is located the upper and lower position of roof, and then the freezing gas of depositing the intracavity in advance gets into the apron through U type hole in, freezes the top of depositing the pipe and deposits, and the catch bar can promote the sliding plate and move down a segment distance in addition, can further open the air vent this moment, and the entering that makes freezing gas in the liquid nitrogen storehouse can be more freezes in depositing chamber and the apron in advance, otherwise the through-hole of U type downthehole portion extends to the roof in, avoids freezing gas of depositing the intracavity in advance to extend to the external world, avoids freezing gas to overflow through U type hole and scatters.

In a possible design, the top fixedly connected with display screen of apron, the top inner wall fixedly connected with temperature sensor of apron, and temperature sensor and display screen electricity are connected, a plurality of rubber kicking blocks of the top inner wall fixedly connected with of apron, and the bottom of rubber kicking block extends to and freezes in the storage sleeve, one side of freezing the storage box is fixed to be run through has the liquid nitrogen filling tube, and the one end of liquid nitrogen filling tube extends to the liquid nitrogen storehouse in, when freezing the storage tube and placing the back that finishes, rotates the apron and seals the freezing storage box, can real-time supervision apron and the temperature of freezing the storage intracavity in advance through temperature sensor, guarantees to freeze gas and can carry out effectual freezing to freezing the storage tube, and the rubber kicking block extends to and freezes in the storage sleeve and carries out preliminary fixed to freezing the storage tube.

In one possible design, one side of the cryopreservation box, which is far away from each other, is provided with a sliding groove, a pin shaft is connected in the sliding groove in a sliding manner, a plurality of freezing air through holes are formed in the fixed plate, two sides of the cryopreservation box are provided with sliding pins in a sliding manner, one ends, which are far away from each other, of the two sliding pins extend into the corresponding sliding grooves respectively, one ends, which are close to each other, of the two sliding pins are fixedly connected with a sliding plate, the sliding plate is connected with the top of the fixed plate in a sliding manner, the sliding plate is internally provided with connecting holes communicated with the freezing air through holes, the outer wall of each sliding pin is fixedly sleeved with a fixed ring, the outer wall of each sliding pin is sleeved with a tension spring fixedly connected with the fixed ring, and one end, which is far away from the fixed ring, of the tension spring is fixedly connected with the inner wall of one side of the cryopreservation box, one side that the round pin axle is close to the cryopreserved box is equipped with the draw-in groove with sliding pin looks block, the draw-in groove is kept away from one side inner wall slip of sliding pin and is run through the push rod, and the one end slip of push rod runs through the handle, the handle drives the round pin axle and moves down along the orbit of sliding tray, the handle is suppressed the apron, the round pin axle promotes the sliding pin and moves to the inboard, and the sliding pin just blocks in the draw-in groove, and then can guarantee the handle can further seal the apron through the braking of sliding pin axle to the round pin axle, the removal of slide can make the connecting hole align with freezing gas through-hole, freezing gas in the liquid nitrogen storehouse can further get into through freezing gas through-hole and connecting hole in the cryopreserved chamber, increase the cryopreserved chamber and cryopreserved pipe and the cryopreserved effect of cryopreserved pipe.

The CAR-T cell cryopreservation box cryopreservation method comprises the following steps:

s1, opening a cover plate, respectively injecting CAR-T cell suspension liquid into a plurality of freezing pipes, sequentially placing the freezing pipes into a freezing sleeve, when the freezing pipes are placed into the freezing sleeve, pushing a sliding rod, a tray, a sealing plate and a transverse plate to move downwards by the gravity of the freezing pipes and the CAR-T cell suspension liquid, aligning a first through hole with a second through hole, further enabling frozen gas emitted by liquid nitrogen in a liquid nitrogen bin to penetrate into a pre-freezing chamber after being reduced by a fixing plate and a sliding plate, enabling the reduced frozen gas in the pre-freezing chamber to enter the freezing sleeve through the first through hole and the second through hole, and primarily freezing the freezing pipes, wherein the sliding plate and the fixing plate are arranged to prevent the frozen gas of the liquid nitrogen in the liquid nitrogen bin from overflowing;

s2, when the number of the freezing pipes placed in the freezing sleeve is increased, the transverse plates are driven by the gravity of the freezing pipes to downwards extrude the sliding plates, the sliding plates start to downwards move for a small distance, the sliding plates extrude the second springs through the second fixing columns, the sliding plates drive the connecting rods to rotate, the connecting rods push the sliding sealing blocks to move for a certain distance, and therefore plugging of the sliding sealing blocks on the vent holes is gradually removed, freezing gas in the liquid nitrogen bin enters the pre-freezing chamber through the vent holes and the round holes in the sliding plates, freezing gas in the pre-freezing chamber is increased, the freezing pipes placed in the prior art are further frozen, namely rapid overflow of the freezing gas in the liquid nitrogen bin is avoided, the prior freezing pipes can be frozen, and deterioration of the prior freezing pipes is avoided;

s3, after the freezing pipe is placed, closing the cover plate, sealing the freezing box by the cover plate, pushing the push rod to move downwards by the cover plate, wherein holes at the bottom of the U-shaped hole are always positioned in the top plate, and after the cover plate pushes the push rod to move downwards, holes at two ends of the U-shaped hole are just positioned at the upper position and the lower position of the top plate, so that the frozen gas in the pre-freezing cavity enters the cover plate through the U-shaped hole to freeze the top of the freezing pipe;

s4, the handle is pushed to move downwards, the handle drives the pin shaft to move downwards along the track of the sliding groove, the inner wall of the top of the handle is attached to the top of the cover plate, the pin shaft pushes the sliding pin and the sliding plate to move inwards, the sliding pin is just clamped into the clamping groove, the handle can be further sealed by the sliding pin to brake the pin shaft, the cover plate is prevented from being opened, in addition, the sliding plate can be moved to enable the connecting hole to be aligned with the freezing gas through hole, the freezing gas in the liquid nitrogen bin can further enter the pre-freezing storage cavity through the freezing gas through hole and the connecting hole, the freezing storage effect of the pre-freezing storage cavity and the freezing storage pipe on the freezing storage pipe is increased, then the freezing storage box is transported until the plurality of freezing storage pipes are transported to corresponding large-scale freezing storage equipment in a unified mode to be frozen and stored for a long time.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.

According to the freezing and storing device, a sliding rod penetrates through the inner wall of the bottom of a freezing and storing sleeve in a sliding mode, a transverse plate is fixedly connected to the bottom end of the sliding rod, a sealing plate is fixedly connected to two sides of the top of the transverse plate, a plurality of first through holes are formed in two sides of the freezing and storing sleeve, a plurality of second through holes matched with the first through holes are formed in the sealing plate, the first through holes and the second through holes are mutually staggered to seal the first through holes under normal conditions, the situation that frozen gas in a pre-freezing and storing cavity enters the freezing and storing sleeve to cause frozen gas overflow and scattering is avoided, when the freezing and storing tube is placed in the freezing and storing sleeve, the sliding rod is pushed by the gravity of a CAR-T cell suspension, the first through holes are aligned with the second through holes, the frozen gas in the pre-freezing and storing cavity enters the freezing and storing sleeve, and the freezing and storing tube is frozen preliminarily;

according to the invention, the top of the fixed plate is slidably connected with a sliding sealing block for sealing the vent hole, the top of the sliding sealing block is rotatably connected with a connecting rod, the top end of the connecting rod is rotatably connected with the bottom of the sliding plate, when the number of the freezing tubes placed in the freezing sleeve is increased, the transverse plate is driven by the gravity of the freezing tubes to downwards extrude the sliding plate, the sliding plate drives the connecting rod to rotate, the connecting rod pushes the sliding sealing block to move for a certain distance, so that the sliding sealing block is gradually removed from sealing the vent hole, the frozen gas in the liquid nitrogen bin enters the pre-freezing chamber, and the freezing tubes placed in advance are further frozen, so that the rapid overflow of the frozen gas in the liquid nitrogen bin is avoided, the freezing tubes in advance can be frozen, and the deterioration of the freezing tubes in advance is avoided;

according to the invention, a plurality of push rods penetrate through the top plate in a sliding manner, the bottom ends of the push rods are fixedly connected with the top of the sliding plate, U-shaped holes are formed in the push rods, the upper through holes and the lower through holes of the U-shaped holes are respectively positioned at the upper position and the lower position of the top plate, after the push rods are pushed to move downwards by the cover plate, the holes at the two ends of the U-shaped holes are just positioned at the upper position and the lower position of the top plate, so that frozen gas in the pre-freezing storage cavity enters the cover plate through the U-shaped holes, the top of the freezing storage tube is frozen, in addition, the push rods push the sliding plate to move downwards for a short distance, and further vent holes are opened, so that more frozen gas in the liquid nitrogen bin can enter the pre-freezing storage cavity and the cover plate, otherwise, the through holes at the bottoms of the U-shaped holes extend into the top plate, the frozen gas in the pre-freezing storage cavity is prevented from extending to the outside, and the frozen gas is prevented from overflowing and scattering through the U-shaped holes;

according to the freezing storage box, sliding pins penetrate through two sides of the freezing storage box in a sliding mode, one ends, close to each other, of the two sliding pins are fixedly connected with sliding plates, one sides, close to the freezing storage box, of the pin shafts are provided with clamping grooves clamped with the sliding pins, the pin shafts are driven by handles to move downwards along the tracks of the sliding grooves, the sliding pins are pushed to move inwards, the sliding pins are just clamped into the clamping grooves, the handles can further seal the cover plate through braking of the sliding pins, the connecting holes can be aligned with the freezing air through holes through the sliding plates in a moving mode, freezing air in the liquid nitrogen bin can further enter the freezing storage cavity through the freezing air through holes and the connecting holes, and the freezing storage effect of the freezing storage cavity and the freezing storage pipe on the freezing storage pipe is improved.

According to the invention, the freezing tube is placed in the freezing sleeve, and the sealing of the sealing plate on the first through hole can be removed through the dead weight of the freezing tube and CAR-T cell suspension liquid, so that the freezing tube can be primarily frozen.

Drawings

FIG. 1 is a schematic diagram of the three-dimensional structure of a CAR-T cell cryopreservation box provided by an embodiment of the invention;

FIG. 2 is a schematic diagram of the three-dimensional structure of the interior of a CAR-T cell cryopreservation box according to an embodiment of the invention;

FIG. 3 is a schematic sectional view of a CAR-T cell cryopreservation box according to an embodiment of the present invention;

FIG. 4 is a schematic diagram of a three-dimensional explosive structure of a CAR-T cell cryopreservation cartridge encasement rack provided by an embodiment of the invention;

FIG. 5 is a schematic three-dimensional cross-sectional view of a cryopreserving sleeve and a closing plate of a CAR-T cell cryopreserving cassette according to an embodiment of the present invention;

FIG. 6 is a schematic diagram of a three-dimensional cross-sectional structure of a first thermal-insulating cylinder of a CAR-T cell cryopreservation box according to an embodiment of the invention;

FIG. 7 is a schematic diagram of a three-dimensional cross-sectional structure of a sliding closure block of a CAR-T cell cryopreservation cassette according to an embodiment of the invention;

FIG. 8 is a schematic diagram of a three-dimensional cross-sectional structure of a buffer structure of a CAR-T cell cryopreservation box according to an embodiment of the invention;

FIG. 9 is a schematic diagram of a three-dimensional cross-sectional structure of a top plate and a push rod of a CAR-T cell cryopreservation box provided by an embodiment of the invention;

FIG. 10 is a schematic sectional view of the cryopreservation box of the CAR-T cell cryopreservation box provided by the embodiment of the invention in the second embodiment;

FIG. 11 is an enlarged schematic structural view of the CAR-T cell cryopreservation box provided by the second embodiment of the invention at position A.

Reference numerals:

1. freezing and storing the box; 2. a cover plate; 3. a pin shaft; 4. a handle; 5. a temperature sensor; 6. a display screen; 7. a rubber top block; 8. a top plate; 9. freezing and storing the sleeve; 10. a sliding plate; 11. a fixing plate; 12. a liquid nitrogen bin; 13. pre-freezing and storing the cavity; 14. a slide bar; 15. a tray; 16. a closing plate; 17. a transverse plate; 18. a first through hole; 19. a second through hole; 20. a first thermal insulation cylinder; 21. a first spring; 22. a first fixed column; 23. a vent hole; 24. sliding the closing block; 25. a connecting rod; 26. a push rod; 27. a U-shaped hole; 28. a buffer structure; 29. a second thermal insulation cylinder; 30. a second spring; 31. a second fixed column; 32. a liquid nitrogen injection pipe; 33. a sliding groove; 34. a slide pin; 35. a fixing ring; 36. a tension spring; 37. a card slot; 38. a push rod; 39. a frozen gas through hole; 40. a slide plate; 41. and connecting the holes.

Detailed Description

The embodiments of the present invention will be described below with reference to the drawings.

In the description of the embodiments of the present invention, it should be noted that, unless explicitly stated or limited otherwise, the terms "connected" and "mounted" are to be interpreted broadly, e.g., "connected" may or may not be detachably connected; may be directly connected or indirectly connected through an intermediate. Further, "communication" may be direct communication or indirect communication through an intermediary. The term "fixed" means that they are connected to each other and the relative positional relationship after the connection is not changed. Directional phrases used in embodiments of the present invention, such as "inner", "outer", "top", "bottom", and the like, refer only to the orientation of the attached drawings and, therefore, are used in order to better and more clearly describe and understand the embodiments of the present invention, and do not indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be taken as limiting the embodiments of the present invention.

In the embodiments of the present invention, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or to implicitly indicate the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature.

In the embodiment of the present invention, "and/or" is only one kind of association relationship describing an association object, and indicates that three relationships may exist, for example, a and/or B may indicate: a exists alone, A and B exist simultaneously, and B exists alone. In addition, the character "/" herein generally indicates that the former and latter related objects are in an "or" relationship.

Reference throughout this specification to "one embodiment" or "some embodiments," or the like, means that a particular feature, structure, or characteristic described in connection with the embodiment is included in one or more embodiments of the present invention. Thus, appearances of the phrases "in one embodiment," "in some embodiments," "in other embodiments," or the like, in various places throughout this specification are not necessarily all referring to the same embodiment, but rather mean "one or more but not all embodiments" unless specifically stated otherwise. The terms "comprising," "including," "having," and variations thereof mean "including, but not limited to," unless otherwise specifically stated.

Example 1

Referring to figures 1, 2, 3 and 4, a CAR-T cell cryopreservation cassette of the present embodiment comprises: cryopreserved box 1, cryopreserved box 1's top hinge has apron 2, cryopreserved box 1's the equal fixedly connected with round pin axle 3 in both sides, the one end that two round pin axles 3 kept away from each other is rotated and is connected with same handle 4, cryopreserved box 1 sliding connection has sliding plate 10, cryopreserved box 1 internal fixation has the fixed plate 11 that is located sliding plate 10 below, and through 28 elastic connection of buffer structure between sliding plate 10 and the fixed plate 11, the structure is deposited in the prefreezing, the setting is in cryopreserved box 1, be used for tentatively cryopreserving the pipe, enlarge and freeze the gas structure, the setting is in fixed plate 11, a volume for increasing and freeze gas, be convenient for freeze and reserve the pipe fast.

Referring to fig. 1, fig. 2 and fig. 3, bolt fixedly connected with display screen 6 is passed through at the top of apron 2, the top inner wall of apron 2 passes through bolt fixedly connected with temperature sensor 5, and temperature sensor 5 is connected with 6 electricity of display screen, a plurality of rubber kicking blocks 7 of bolt fixedly connected with are passed through to the top inner wall of apron 2, and the bottom of rubber kicking block 7 extends to in the cryopreserved sleeve 9, one side of cryopreserved box 1 is fixed to be run through there is liquid nitrogen injection pipe 32, and the one end of liquid nitrogen injection pipe 32 extends to in the liquid nitrogen storehouse 12, after the cryopreserved pipe is placed and finishes, it freezes the box 1 to rotate apron 2 and seal, can real-time supervision apron 2 and the temperature in the prefreezing reserved chamber 13 through temperature sensor 5, guarantee that the gas of freezing can carry out effectual freezing to the cryopreserved pipe, and rubber kicking block 7 extends to and freezes in the cryopreserved sleeve 9 and carry out preliminary fixed to the cryopreserved pipe.

Referring to fig. 5 and 6, the pre-freezing structure includes a top plate 8 fixedly connected in a freezing box 1 through bolts, a plurality of freezing sleeves 9 are fixedly penetrated in the top plate 8, sliding rods 14 are slidably penetrated through the inner walls of the bottoms of the freezing sleeves 9, the top ends of the sliding rods 14 are fixedly connected with trays 15 positioned in the freezing sleeves 9 through bolts, the bottom ends of the sliding rods 14 are fixedly connected with a transverse plate 17 through bolts, both sides of the tops of the transverse plate 17 are fixedly connected with a sealing plate 16 through bolts, both sides of the freezing sleeves 9 are provided with a plurality of first through holes 18, a plurality of second through holes 19 matched with the first through holes 18 are arranged in the sealing plate 16, under normal conditions, the first through holes 18 and the second through holes 19 are staggered with each other, the first through holes 18 can be sealed by the sealing plate 16, frozen gas in the pre-freezing chamber 13 is prevented from entering the freezing sleeves 9 to cause frozen gas overflow, when the freezing tube is put into the freezing sleeve 9, gravity of the freezing tube pushes the-T cell suspension to penetrate into the sliding rods 14 and the sealing plate 16, the sliding rods move downwards, and then the freezing chamber 18 is aligned with the freezing chamber 12 to absorb the frozen liquid nitrogen before freezing tube 11.

Referring to fig. 6, the top of the sliding plate 10 is fixedly connected with a plurality of first heat insulation cylinders 20 through bolts, the inner wall of the bottom of each first heat insulation cylinder 20 is fixedly connected with a first spring 21, each first heat insulation cylinder 20 is slidably connected with a first fixing column 22 fixedly connected with the top end of the corresponding first spring 21, the top end of each first fixing column 22 is fixedly connected with the bottom of the corresponding transverse plate 17 through bolts, and through the cooperation of the corresponding first fixing column 22 and the corresponding first fixing column 22, the sealing of the corresponding first through hole 18 by the sealing plate 16 can be released according to the gravity of the cryopreserved pipe and the CAR-T cell suspension, so that the phenomenon that the frozen gas overflows through the corresponding first through hole 18 can be avoided, and the cryopreserved pipe can be frozen in time.

Referring to fig. 7, the structure for expanding frozen gas comprises a plurality of vent holes 23 arranged in a fixed plate 11, the top of the fixed plate 11 is slidably connected with a sliding closed block 24 for closing the vent holes 23, the top of the sliding closed block 24 is rotatably connected with a connecting rod 25, and the top end of the connecting rod 25 is rotatably connected with the bottom of the sliding plate 10, when the number of the frozen pipes put into the freezing sleeve 9 increases, the gravity of the plurality of frozen pipes drives the transverse plate 17 to downwardly extrude the sliding plate 10, the sliding plate 10 downwardly moves a small distance, the sliding plate 10 drives the connecting rod 25 to rotate, the connecting rod 25 pushes the sliding closed block 24 to move a certain distance, so as to gradually remove the blocking of the sliding closed block 24 on the vent holes 23, frozen gas in a liquid nitrogen bin 12 enters a pre-freezing storage chamber 13 through the vent holes 23 and round holes on the sliding plate 10, frozen gas in the pre-freezing chamber 13 is increased, the frozen pipes placed previously are further frozen, namely, rapid overflow and scattering of the frozen gas in the liquid nitrogen bin 12 is avoided, and the frozen pipes previously can be frozen.

Referring to fig. 4 and 8, the buffer structure 28 includes a plurality of second thermal insulation cylinders 29 fixedly connected to the top of the fixed plate 11 by bolts, a second spring 30 is fixedly connected to the inner wall of the bottom of the second thermal insulation cylinders 29, the elastic force of the second spring 30 is greater than the elastic force of the first spring 21, second fixing columns 31 fixedly connected to the top ends of the second springs 30 are slidably connected to the second thermal insulation cylinders 29, the top ends of the second fixing columns 31 are fixedly connected to the bottom of the sliding plate 10 by bolts, the sliding plate 10 can be prevented from moving downwards when the number of the cryopreserved pipes is small by the cooperation of the second spring 30 and the second fixing columns 31, the vent holes 23 are prevented from being opened, when the number of the cryopreserved pipes is relatively small, the sliding plate 10 can push the second fixing columns 31 to move downwards, the vent holes 23 are gradually opened, and further more frozen gas can enter the prefreefer chamber 13 to freeze the cryopreserved pipes.

Refer to 3, fixed plate 11 forms liquid nitrogen storehouse 12 with the bottom inner wall that freezes and deposits box 1, the top of sliding plate 10 forms with the bottom of roof 8 and freezes in advance and deposits chamber 13, and the bottom that freezes to deposit sleeve 9 extends to in freezing in advance and deposits chamber 13, be equipped with a plurality of round holes in the sliding plate 10, freezing gas in the liquid nitrogen storehouse 12 permeates the back penetration after the heat preservation of sliding plate 10 and fixed plate 11 and freezes in advance and deposits chamber 13, just in freezing in advance and depositing chamber 13 remaining freezing gas can tentatively freeze the pipe and deposit, can avoid freezing gas in the liquid nitrogen storehouse 12 to overflow and scatter again.

Referring to fig. 9, a plurality of push rods 26 are slidably penetrated in the top plate 8, the bottom ends of the push rods 26 are fixedly connected with the top of the sliding plate 10 through bolts, U-shaped holes 27 are formed in the push rods 26, upper and lower through holes of the U-shaped holes 27 are respectively located at upper and lower positions of the top plate 8, the cover plate 2 pushes the push rods 26 to move downwards, holes at two ends of the U-shaped holes 27 are just located at upper and lower positions of the top plate 8, frozen gas in the pre-frozen storage cavity 13 enters the cover plate 2 through the U-shaped holes 27, the top of a frozen storage pipe is frozen, the push rods 26 can push the sliding plate 10 to move downwards for a short distance, at the moment, the vent holes 23 can be further opened, frozen gas in the liquid nitrogen bin 12 can enter the pre-frozen storage cavity 13 and the cover plate 2 more, otherwise, the through holes at the bottoms of the U-shaped holes 27 extend into the top plate 8, the frozen gas in the pre-frozen storage cavity 13 is prevented from extending to the outside, and the frozen gas is prevented from overflowing through the U-shaped holes 27.

Example 2

Referring to fig. 6, the top of the sliding plate 10 is fixedly connected with a plurality of first heat insulation cylinders 20 through bolts, the inner wall of the bottom of each first heat insulation cylinder 20 is fixedly connected with a first spring 21, the first heat insulation cylinders 20 are slidably connected with first fixing columns 22 fixedly connected with the top ends of the first springs 21, the top ends of the first fixing columns 22 are fixedly connected with the bottom of the transverse plate 17 through bolts, and through the cooperation of the first fixing columns 22 and the first fixing columns 22, the sealing of the first through holes 18 by the sealing plate 16 can be released according to the gravity of the cryopreservation tubes and the CAR-T cell suspension, so that the cryopreservation tubes can be prevented from overflowing and scattering through the first through holes 18, and can be frozen in time.

Referring to fig. 7, the structure for expanding frozen gas includes a plurality of vent holes 23 arranged in a fixed plate 11, the top of the fixed plate 11 is slidably connected with a sliding sealing block 24 for sealing the vent holes 23, the top of the sliding sealing block 24 is rotatably connected with a connecting rod 25, and the top end of the connecting rod 25 is rotatably connected with the bottom of the sliding plate 10, when the number of the frozen pipes put into the frozen sleeve 9 increases, the gravity of the frozen pipes drives the transverse plate 17 to downwards extrude the sliding plate 10, the sliding plate 10 downwards moves a small distance, the sliding plate 10 drives the connecting rod 25 to rotate, the connecting rod 25 pushes the sliding sealing block 24 to move a certain distance, so as to gradually remove the blockage of the sliding sealing block 24 on the vent holes 23, frozen gas in a liquid nitrogen bin 12 enters the pre-frozen storage cavity 13 through the vent holes 23 and round holes on the sliding plate 10, frozen gas in the pre-frozen storage cavity 13 is increased, further frozen pipes put in advance are frozen, that the frozen pipes are put in advance are prevented from being quickly overflowed by the frozen gas in the liquid nitrogen bin 12, and the frozen pipes are prevented from being frozen and deteriorated.

Refer to 3, fixed plate 11 forms liquid nitrogen storehouse 12 with the bottom inner wall that freezes deposits box 1, the top of sliding plate 10 forms with the bottom of roof 8 and freezes to deposit chamber 13 in advance, and the bottom that freezes deposits sleeve 9 extends to in freezing in advance and deposits chamber 13, be equipped with a plurality of round holes in the sliding plate 10, freezing gas in the liquid nitrogen storehouse 12 passes through sliding plate 10 and fixed plate 11's heat preservation after the infiltration in freezing in advance and deposits chamber 13, just freezing remaining freezing gas in depositing chamber 13 in advance can tentatively freeze the pipe and deposit, can avoid freezing gas in the liquid nitrogen storehouse 12 to overflow and scatter again.

Referring to fig. 10 and 11, the freezing storage box 1 is provided with a sliding groove 33 at one side far from each other, the pin shaft 3 is slidably connected in the sliding groove 33, the fixing plate 11 is provided with a plurality of freezing air through holes 39, the two sides of the freezing storage box 1 are provided with sliding pins 34 in a sliding way, the ends of the two sliding pins 34 far from each other respectively extend into the corresponding sliding grooves 33, the ends of the two sliding pins 34 close to each other are fixedly connected with a sliding plate 40 through bolts, the sliding plate 40 is slidably connected with the top of the fixing plate 11, the sliding plate 40 is provided with a connecting hole 41 communicated with the freezing air through hole 39, the outer wall of the sliding pin 34 is fixedly sleeved with a fixing ring 35, the outer wall of the sliding pin 34 is sleeved with a tension spring 36 fixedly connected with the fixing ring 35, and the end of the tension spring 36 far from the fixing ring 35 is fixedly connected with the inner wall of one side of the freezing storage box 1, one side of the pin shaft 3 close to the cryopreservation box 1 is provided with a clamping groove 37 clamped with the sliding pin 34, the inner wall of one side of the clamping groove 37 far away from the sliding pin 34 slides to penetrate through a push rod 38, one end of the push rod 38 slides to penetrate through the handle 4, the handle 4 drives the pin shaft 3 to move downwards along the track of the sliding groove 33, the handle 4 presses the cover plate 2, the pin shaft 3 pushes the sliding pin 34 to move inwards, the sliding pin 34 is just clamped into the clamping groove 37, and then the handle 4 can be further sealed by braking the sliding pin 34 on the pin shaft 3, the connecting hole 41 can be aligned with the freezing gas through hole 39 through the movement of the sliding plate 40, the freezing gas in the liquid nitrogen bin 12 can further enter the pre-cryopreservation cavity 13 through the freezing gas through hole 39 and the connecting hole 41, and the cryopreservation effect of the pre-cryopreservation cavity 13 and the cryopreservation tube is improved.

A method of cryopreserving a CAR-T cell cryopreserving cassette, comprising the steps of:

s1, opening a cover plate 2, respectively injecting CAR-T cell suspension liquid into a plurality of freezing tubes, sequentially placing the freezing tubes into a freezing sleeve 9, when the freezing tubes are placed into the freezing sleeve 9, pushing a sliding rod 14, a tray 15, a sealing plate 16 and a transverse plate 17 to move downwards by the gravity of the freezing tubes and the CAR-T cell suspension liquid, aligning a first through hole 18 with a second through hole 19, further leading the frozen gas diffused by liquid nitrogen in a liquid nitrogen bin 12 to penetrate into a pre-freezing chamber 13 after being reduced by a fixing plate 11 and a sliding plate 10, leading the reduced frozen gas in the pre-freezing chamber 13 to enter the freezing sleeve 9 through the first through hole 18 and the second through hole 19, primarily freezing the freezing tubes, and avoiding the frozen gas of the liquid nitrogen in the liquid nitrogen bin 12 from overflowing and scattering due to the arrangement of the sliding plate 10 and the fixing plate 11;

s2, when the number of the freezing pipes placed in the freezing sleeve 9 is increased, the gravity of the freezing pipes drives the transverse plate 17 to downwards extrude the sliding plate 10, the sliding plate 10 starts to downwards move for a short distance, the sliding plate 10 extrudes the second spring 30 through the second fixing column 31, the sliding plate 10 drives the connecting rod 25 to rotate, the connecting rod 25 pushes the sliding sealing block 24 to move for a certain distance, so that the sliding sealing block 24 is gradually removed from blocking the vent hole 23, the frozen gas in the liquid nitrogen bin 12 enters the pre-freezing chamber 13 through the vent hole 23 and the round holes in the sliding plate 10, the frozen gas in the pre-freezing chamber 13 is increased, and the freezing pipes placed previously are further frozen, so that the frozen gas in the liquid nitrogen bin 12 is prevented from quickly overflowing, the previous freezing pipes can be frozen, and the previous freezing pipes are prevented from deteriorating;

s3, after the freezing storage pipe is placed, closing the cover plate 2, sealing the freezing storage box 1 by the cover plate 2, pushing the push rod 26 to move downwards by the cover plate 2, wherein the hole at the bottom of the U-shaped hole 27 is always positioned in the top plate 8, after the push rod 26 is pushed to move downwards by the cover plate 2, the holes at two ends of the U-shaped hole 27 are just positioned at the upper position and the lower position of the top plate 8, so that the freezing gas in the pre-freezing storage cavity 13 enters the cover plate 2 through the U-shaped hole 27 to freeze the top of the freezing storage pipe, in addition, the push rod 26 can push the sliding plate 10 to move downwards for a small distance, and at the moment, the vent hole 23 can be further opened, so that more freezing gas in the liquid nitrogen bin 12 can enter the pre-freezing storage cavity 13 and the cover plate 2;

s4, the handle 4 is pushed to move downwards, the handle 4 drives the pin shaft 3 to move downwards along the track of the sliding groove 33, the inner wall of the top of the handle 4 is attached to the top of the cover plate 2, the pin shaft 3 pushes the sliding pin 34 and the sliding plate 40 to move inwards, the sliding pin 34 is just clamped into the clamping groove 37, further the handle 4 can be guaranteed to further seal the cover plate 2 through the braking of the sliding pin 34 on the pin shaft 3, the cover plate 2 is prevented from being opened, in addition, the connecting hole 41 can be aligned to the freezing through hole 39 through the movement of the sliding plate 40, freezing gas in the liquid nitrogen bin 12 can further enter the pre-freezing chamber 13 through the freezing through hole 39 and the connecting hole 41, the freezing effect of the pre-freezing chamber 13 and the freezing pipe on the freezing pipe is increased, the freezing box 1 is transported, and the freezing pipes are transported uniformly to corresponding large-scale freezing equipment for long-term freezing.

However, as is well known to those skilled in the art, the working principle and wiring method of the temperature sensor 5 and the display screen 6 are common and are conventional means or common knowledge, and thus will not be described in detail herein, and those skilled in the art can make any choice according to their needs or convenience.

The above description is only an embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art can easily think of the changes or substitutions within the technical scope of the present invention, and the changes or substitutions should be covered within the scope of the present invention; the embodiments of the invention and the features of the embodiments can be combined with each other without conflict. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims

1. A CAR-T cell cryopreservation cassette comprising:

the freezing storage box (1) is hinged with a cover plate (2) at the top of the freezing storage box (1), pin shafts (3) are fixedly connected to two sides of the freezing storage box (1), and one end, far away from each other, of each of the two pin shafts (3) is rotatably connected with the same handle (4);

a sliding plate (10) is connected in the freezing box (1) in a sliding manner, a fixed plate (11) positioned below the sliding plate (10) is fixedly connected in the freezing box (1), and the sliding plate (10) is elastically connected with the fixed plate (11) through a buffer structure (28);

the pre-freezing structure is arranged in the freezing storage box (1) and is used for primarily freezing the freezing storage tube;

the freezing gas expansion structure is arranged in the fixing plate (11) and used for increasing the amount of freezing gas and facilitating quick freezing of the freezing storage pipe.

2. A CAR-T cell cryopreservation box as claimed in claim 1, wherein the pre-cryopreservation structure comprises a top plate (8) fixedly connected in the cryopreservation box (1), a plurality of cryopreservation sleeves (9) are fixedly arranged in the top plate (8), a sliding rod (14) is slidably arranged on the inner wall of the bottom of each cryopreservation sleeve (9), a tray (15) positioned in each cryopreservation sleeve (9) is fixedly connected to the top end of each sliding rod (14), a transverse plate (17) is fixedly connected to the bottom end of each sliding rod (14), a sealing plate (16) is fixedly connected to both sides of the top of each transverse plate (17), a plurality of first through holes (18) are arranged on both sides of each cryopreservation sleeve (9), and a plurality of second through holes (19) matched with the first through holes (18) are arranged in each sealing plate (16).

3. A CAR-T cell cryopreservation cartridge (1) according to claim 1, wherein the expanding freezing gas structure comprises a plurality of vent holes (23) arranged in a fixed plate (11), the top of the fixed plate (11) is slidably connected with a sliding closure block (24) for closing the vent holes (23), the top of the sliding closure block (24) is rotatably connected with a connecting rod (25), and the top end of the connecting rod (25) is rotatably connected with the bottom of the sliding plate (10).

4. A CAR-T cell cryopreservation box as claimed in claim 1, wherein a plurality of first thermal insulation cylinders (20) are fixedly connected to the top of the sliding plate (10), a first spring (21) is fixedly connected to the inner wall of the bottom of the first thermal insulation cylinders (20), a first fixing column (22) fixedly connected to the top end of the first spring (21) is slidably connected to the first thermal insulation cylinders (20), and the top end of the first fixing column (22) is fixedly connected to the bottom of the transverse plate (17).

5. A CAR-T cell cryopreservation cartridge as claimed in claim 1, wherein the buffer structure (28) comprises a plurality of second temperature-insulating cylinders (29) fixedly connected to the top of the fixed plate (11), the inner walls of the bottoms of the second temperature-insulating cylinders (29) are fixedly connected with second springs (30), the elastic force of the second springs (30) is greater than that of the first springs (21), second fixing columns (31) fixedly connected to the top ends of the second springs (30) are slidably connected to the second temperature-insulating cylinders (29), and the top ends of the second fixing columns (31) are fixedly connected to the bottom of the sliding plate (10).

6. A CAR-T cell cryopreservation box according to claim 1, characterized in that the fixed plate (11) and the bottom inner wall of the cryopreservation box (1) form a liquid nitrogen bin (12), the top of the sliding plate (10) and the bottom of the top plate (8) form a pre-cryopreservation cavity (13), the bottom end of the cryopreservation sleeve (9) extends into the pre-cryopreservation cavity (13), and the sliding plate (10) is provided with a plurality of round holes therein.

7. A CAR-T cell cryopreservation box as claimed in claim 2, wherein a plurality of push rods (26) are slidably inserted in the top plate (8), the bottom ends of the push rods (26) are fixedly connected with the top of the sliding plate (10), U-shaped holes (27) are formed in the push rods (26), and the upper and lower through holes of the U-shaped holes (27) are respectively located at the upper and lower positions of the top plate (8).

8. The CAR-T cell cryopreservation box of claim 1, wherein a display screen (6) is fixedly connected to the top of the cover plate (2), a temperature sensor (5) is fixedly connected to the inner wall of the top of the cover plate (2), the temperature sensor (5) is electrically connected with the display screen (6), a plurality of rubber top blocks (7) are fixedly connected to the inner wall of the top of the cover plate (2), the bottom ends of the rubber top blocks (7) extend into the cryopreservation sleeve (9), a liquid nitrogen injection pipe (32) is fixedly penetrated through one side of the cryopreservation box (1), and one end of the liquid nitrogen injection pipe (32) extends into the liquid nitrogen bin (12).

9. A CAR-T cell cryopreservation box according to claim 1, wherein the cryopreservation box (1) is provided with a sliding groove (33) at one side away from each other, a pin shaft (3) is slidably connected in the sliding groove (33), a plurality of freezing through holes (39) are arranged in the fixing plate (11), sliding pins (34) penetrate through the two sides of the cryopreservation box (1), two ends of the sliding pins (34) which are away from each other extend into the corresponding sliding grooves (33), two sliding pins (34) are fixedly connected with a sliding plate (40) at one end close to each other, the sliding plate (40) is slidably connected with the top of the fixing plate (11), a connecting hole (41) communicated with the freezing through holes (39) is arranged in the sliding plate (40), a fixing ring (35) is arranged on the outer wall of the sliding pins (34), a tension spring (36) fixedly connected with a clamping groove (35) is arranged on the outer wall of the sliding pins (34), one end of the tension spring (36) far away from the fixing ring (35) is fixedly connected with one side of the cryopreservation the freezing box (1), a clamping pin shaft (34) is connected with a push rod (37), and a clamping pin shaft (34) is arranged on one side close to the sliding pin shaft (34) which is connected with a sliding pin shaft (37), and one end of the push rod (38) penetrates through the handle (4) in a sliding manner.

10. A method of cryopreserving a CAR-T cell cryopreserved cassette according to any one of claims 1 to 9, comprising the steps of:

s1, a cover plate (2) is opened, CAR-T cell suspension liquid is respectively injected into a plurality of freezing tubes, the freezing tubes are sequentially placed into a freezing sleeve (9), when the freezing tubes are placed into the freezing sleeve (9), the gravity of the freezing tubes and the CAR-T cell suspension liquid pushes a sliding rod (14), a tray (15), a sealing plate (16) and a transverse plate (17) to move downwards, at the moment, a first through hole (18) is aligned with a second through hole (19), further, frozen gas diffused by liquid nitrogen in a liquid nitrogen bin (12) penetrates into a pre-freezing storage cavity (13) after being reduced through a fixing plate (11) and a sliding plate (10), the reduced frozen gas in the pre-freezing storage cavity (13) enters the freezing sleeve (9) through the first through hole (18) and the second through hole (19), the freezing tubes are frozen preliminarily, and the liquid nitrogen in the freezing tubes can be prevented from overflowing due to the arrangement of the sliding plate (10) and the fixing plate (11);

s2, when the number of the freezing pipes placed in the freezing sleeve (9) is increased, the gravity of the freezing pipes drives the transverse plate (17) to downwards extrude the sliding plate (10), the sliding plate (10) starts to downwards move for a short distance, the sliding plate (10) extrudes the second spring (30) through the second fixing column (31), the sliding plate (10) drives the connecting rod (25) to rotate, the connecting rod (25) pushes the sliding sealing block (24) to move for a certain distance, so that the sliding sealing block (24) can gradually remove the blockage of the vent hole (23), the freezing gas in the liquid nitrogen bin (12) enters the pre-freezing chamber (13) through the vent hole (23) and the circular holes in the sliding plate (10), the freezing gas in the pre-freezing chamber (13) is increased, the freezing pipes placed in advance can be further frozen, namely, the rapid overflow of the freezing gas in the liquid nitrogen bin (12) can be avoided, the freezing of the previous freezing pipes can be also avoided, and the deterioration of the previous freezing pipes can be avoided;

s3, after the frozen pipes are placed, closing a cover plate (2), sealing a frozen box (1) by the cover plate (2), pushing a push rod (26) to move downwards by the cover plate (2), wherein holes at two ends of a U-shaped hole (27) are just positioned at the upper and lower positions of a top plate (8) because the hole at the bottom of the U-shaped hole (27) is always positioned in the top plate (8), and then frozen gas in the pre-frozen storage cavity (13) enters the cover plate (2) through the U-shaped hole (27) to freeze the top of the frozen pipes, and in addition, the push rod (26) can push a sliding plate (10) to move downwards for a small distance, at the moment, a vent hole (23) can be further opened, so that more frozen gas in a liquid nitrogen bin (12) can enter the pre-frozen storage cavity (13) and the cover plate (2);

s4, then, the handle (4) is pushed to move downwards, the handle (4) drives the pin shaft (3) to move downwards along the track of the sliding groove (33), the inner wall of the top of the handle (4) is attached to the top of the cover plate (2), the pin shaft (3) pushes the sliding pin (34) and the sliding plate (40) to move inwards, the sliding pin (34) is just clamped into the clamping groove (37), the cover plate (2) can be further sealed by the handle (4) through the sliding pin (34) to brake the pin shaft (3), the cover plate (2) is prevented from being opened, in addition, the connecting hole (41) can be aligned to the freezing gas through hole (39) through the movement of the sliding plate (40), the freezing gas in the liquid nitrogen bin (12) can further enter the pre-freezing chamber (13) through the freezing gas through hole (39) and the connecting hole (41), the freezing effect of the pre-freezing chamber (13) and the freezing pipe on the freezing pipe is increased, then, the freezing box (1) is transported until a plurality of freezing pipes are transported to corresponding large-scale freezing storage equipment uniformly for a long time.