A sealed adherent stem cell culture factory

Abstract

The application discloses a sealed adherent stem cell culture factory, which comprises a bottom box, a cover body and at least one box body; wherein: adjacent two corners of the box body are respectively provided with an air inlet channel and an air outlet channel, arc-shaped surrounding plates are vertically arranged along edges of the air inlet channel and the air outlet channel, and side holes are formed above the arc-shaped surrounding plates; adjacent two corners of the cover body are respectively provided with an air inlet pipe and an air outlet pipe which are vertically arranged; inner side walls of the bottom box, the box body and the cover body are respectively provided with a plurality of corresponding sealing mechanisms, the bottom of the sealing mechanism is provided with a movable inverted trapezoidal buckle, the top of the sealing mechanism is provided with a buckle hole matched with the inverted trapezoidal buckle, and the bottom box, the box body and the cover body which are adjacently arranged are connected through the sealing mechanism buckles. The sealed adherent stem cell culture factory is flexible and convenient to use, simple to operate, good in connection sealing property and high in stability, can effectively improve the yield of single batch and is suitable for large-scale production in a factory.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biotechnology, in particular to a sealed adherent stem cell culture factory. BACKGROUND

[0002] Cell factory is a kind of cell culture device, which uses the maximum culture surface in limited space, thereby saving space; it can be used for industrial scale production such as vaccine, monoclonal antibody or biopharmaceutical, and is particularly suitable for adherent cells, and can also be used for suspension culture, and when scaling up from laboratory scale, the kinetic conditions of cell growth are not changed, so that scaling up is simple and easy, the risk of contamination is low, and space is saved.

[0003] As disclosed in the published patent CN110283725A, a novel multi-layer adherent cell culture container structure is disclosed, which comprises a multi-layer cell factory main body, a bayonet cover and a gas permeable cover, a plurality of groups of multi-layer culture surfaces are arranged at equal intervals from top to bottom inside the multi-layer cell factory main body, a plurality of multi-layer culture surface fixing grooves are arranged at the multi-layer culture surfaces of the multi-layer cell factory main body, an extension pipe inlet is formed at one side of the upper end of the multi-layer cell factory main body near the base surface and the back surface, the bayonet cover is fixedly installed at the upper end of the extension pipe inlet near the base surface at one side of the upper end of the multi-layer cell factory main body, the gas permeable cover is fixedly installed at the upper end of the extension pipe inlet near the back surface at one side of the upper end of the multi-layer cell factory main body, an extension pipe is arranged inside the gas permeable cover, and a gas permeable film is arranged outside the extension pipe. The multi-layer adherent cell culture container is used for full liquid culture, and although the culture area is greatly increased and the space utilization is improved at the same height, the multi-layer structure is an integral structure, and single-layer or multi-layer cannot be selected for cell culture according to needs, which is not flexible and convenient to use, and resources are wasted to some extent.

[0004] As disclosed in the published patent CN201999918U, a stacked sealed cell culture box is disclosed, which has a rectangular box body, the upper opening of the box body has a wide platform, a step groove is formed along the inner opening of the platform, the outer opening size of the step groove is equal to or greater than the outer shape size of the bottom of the box body, and the inner opening size of the step groove is smaller than the size of the bottom of the box body; an air inlet pipe and an air outlet pipe are arranged vertically in the box body. The stacked sealed cell culture box solves the problems of small culture area, poor sealing, easy contamination, small single culture yield and large space occupation of the existing culture device to some extent, but the sealing method adopted is to set a wide platform at the upper opening of the box body, form a step groove along the inner opening of the platform, and embed the bottom of one box in the platform of the step groove of another box to realize the sealing of the upper and lower two boxes, which has the defects of poor sealing and unstable structure, and is not suitable for the culture and expansion of stem cells.

[0005] For example, the published patent CN109251859A discloses a cell culture device and a cell box thereof. The cell box is a cuboid, comprising a bottom cover, a plurality of partitions and a top cover which are stacked in sequence. The two faces of each partition have a first pool groove and a first rib. The two first pool grooves form a middle layer chamber, and the two first ribs are in close contact to make the multiple middle layer chambers in the same layer independent of each other. The second pool groove of the bottom cover and the first pool groove of the partition form a bottom layer chamber, and the third pool groove of the top cover and the first pool groove of the partition form a top layer chamber. The cell culture device also has the defects of poor sealing and unstable structure, and is not suitable for the culture and expansion of stem cells, which limits its wide use.

[0006] Therefore, how to provide a cell culture device with small pollution risk, high convenience of use, good sealing performance, stable structure, and the ability to culture and harvest a large number of cells at one time and realize automatic blow beating has become a technical problem to be solved by those skilled in the art. SUMMARY

[0007] The technical problem to be solved by the present application is to provide a sealed adherent stem cell culture factory with small pollution risk, high convenience of use, good sealing performance, stable structure, and the ability to culture and harvest a large number of cells at one time.

[0008] To achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0009] The present application provides a sealed adherent stem cell culture factory, comprising a bottom box, a cover body and at least one box body stacked between the bottom box and the cover body; wherein:

[0010] Adjacent two corners of the box body are respectively provided with an air inlet channel and an air outlet channel, and an arc-shaped surrounding plate is vertically arranged along the edges of the air inlet channel and the air outlet channel, and a side hole is formed above the arc-shaped surrounding plate, which communicates the culture area of the box body with the air inlet channel and the air outlet channel;

[0011] Adjacent two corners of the cover body are respectively provided with an air inlet pipe and an air outlet pipe which penetrate up and down, and the air inlet pipe and the air outlet pipe are respectively arranged one by one corresponding to the air inlet channel and the air outlet channel below them;

[0012] The inner side walls of the bottom box, the box body and the cover body are respectively provided with a plurality of corresponding sealing mechanisms, the bottom of the sealing mechanism is provided with a movable inverted trapezoidal buckle, the top is provided with a buckle hole matched with the inverted trapezoidal buckle, and the bottom box, the box body and the cover body arranged adjacent to each other are connected through the sealing mechanism buckle.

[0013] Further, in the sealed adherent stem cell culture factory, each sealing mechanism on the box body and / or the cover body comprises:

[0014] a sealing column arranged on the inner side wall of the box body and / or the cover body, a hollow accommodating chamber being arranged in the sealing column, and a top and a bottom of the sealing column being respectively provided with a bayonet that communicates with the accommodating chamber;

[0015] a T-shaped ejector rod vertically arranged in the accommodating chamber, an annular upper tooth column movably sleeved on an upper end of the T-shaped ejector rod, an annular lower tooth column arranged on an inner wall of the accommodating chamber and matched with the upper tooth column, and a conical tip arranged on a bottom end of the T-shaped ejector rod, the T-shaped ejector rod, the upper tooth column and the lower tooth column forming a rotary telescopic mechanism; and

[0016] an elastic buckle arranged in the accommodating chamber and below the conical tip, the elastic buckle being composed of an inverted L-shaped striding rod, an inverted trapezoidal buckle and a return spring, the inverted trapezoidal buckle being arranged at a lower end bayonet of the accommodating chamber and having two ends respectively connected with the inverted L-shaped striding rod, and the inverted L-shaped striding rod having an upper end arranged in the accommodating chamber through the return spring;

[0017] wherein the rotary telescopic mechanism composed of the T-shaped ejector rod, the upper tooth column and the lower tooth column drives the T-shaped ejector rod to ascend or descend, and the T-shaped ejector rod spreads outward by the conical tip on the bottom end thereof while pushing the inverted L-shaped striding rod and the inverted trapezoidal buckle to descend, so that the inverted trapezoidal buckle is buckled in the top bayonet of another sealing column below the inverted trapezoidal buckle.

[0018] Further preferably, on the sealing type wall-sticking stem cell culture factory, a limiting ring is fixedly arranged on a middle portion of the T-shaped ejector rod, and the upper tooth column is arranged on the limiting ring.

[0019] Further preferably, on the sealing type wall-sticking stem cell culture factory, an inner side of a lower end of the conical tip is recessed with a curved surface, and the curved surface is embedded between the two L-shaped striding rods.

[0020] Further, on the sealing type wall-sticking stem cell culture factory, each of the sealing mechanisms on the bottom box comprises:

[0021] a sealing column arranged on the inner side wall of the bottom box, a hollow accommodating chamber being arranged in the sealing column, and a top and a bottom of the sealing column being respectively provided with a bayonet that communicates with the accommodating chamber; and

[0022] a T-shaped ejector rod vertically arranged in the accommodating chamber, the T-shaped ejector rod abutting at the bayonet at a top of the accommodating chamber through a return spring arranged at a bottom of the accommodating chamber.

[0023] Further, the sealing type adherent stem cell culture factory further comprises a gas permeable cover detachably arranged on the air inlet pipe and the air outlet pipe respectively, the top wall of the gas permeable cover is provided with an opening and a gas permeable film is arranged in the opening.

[0024] Further, the sealing type adherent stem cell culture factory further comprises a side frame arranged at the periphery of the cover body, and a plurality of triangular connecting pieces are arranged at the connection between the top of the cover body and the side frame.

[0025] Further, the sealing type adherent stem cell culture factory further comprises a plurality of reinforcing ribs arranged in parallel along the length direction of the top of the cover body, and a scale line is arranged on the top of the cover body perpendicularly to the reinforcing ribs.

[0026] Further, the sealing type adherent stem cell culture factory further comprises a sealing ring arranged at the bottom periphery of the box body and the cover body respectively.

[0027] Further, the sealing type adherent stem cell culture factory further comprises a pressing cap matched with the T-shaped ejector pin of the sealing mechanism.

[0028] The technical scheme of the present application has the following technical effects compared with the prior art:

[0029] (1) The sealing type adherent stem cell culture factory is a laminated structure composed of the box body arranged between the bottom box and the cover body, which is suitable for the culture of adherent stem cells and ensures the culture area required in the culture process.

[0030] (2) The number of layers of the sealing type adherent stem cell culture factory can be adjusted by controlling the number of stacked box bodies, which is suitable for the culture of different amounts of adherent cells and is flexible and convenient to use.

[0031] (3) The bottom box, the box body and the cover body arranged in layers are connected by the sealing mechanism, which is simple to operate, has good sealing stability, prevents pollution, improves the yield of single batch and is suitable for large-scale production in a factory. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1 Fig. 1 is a schematic view of the overall structure of the sealing type adherent stem cell culture factory of the present application;

[0033] Figure 2 Fig. 2 is a schematic view of the top structure of the bottom box in the sealing type adherent stem cell culture factory of the present application;

[0034] Figure 3 Fig. 3 is a schematic view of the cross-sectional structure of the sealing mechanism on the bottom box in the sealing type adherent stem cell culture factory of the present application;

[0035] Figure 4 It is a top view structural schematic diagram of a box in a sealed adherent stem cell culture factory of the present application;

[0036] Figure 5 It is a bottom view structural schematic diagram of a box in a sealed adherent stem cell culture factory of the present application;

[0037] Figure 6 It is a structural schematic diagram of a box in a sealed adherent stem cell culture factory of the present application before the sealing mechanism buckle;

[0038] Figure 7 It is a structural schematic diagram of a box in a sealed adherent stem cell culture factory of the present application after the sealing mechanism buckle;

[0039] Figure 8 It is a structural schematic diagram of an upper tooth column in a rotary telescopic mechanism of a sealed adherent stem cell culture factory of the present application;

[0040] Figure 9 It is a side view structural schematic diagram of a lower tooth column in a rotary telescopic mechanism of a sealed adherent stem cell culture factory of the present application;

[0041] Figure 10 It is a bottom view structural schematic diagram of a lower tooth column in a rotary telescopic mechanism of a sealed adherent stem cell culture factory of the present application;

[0042] Figure 11 It is a buckle structure schematic diagram of a sealing structure in two adjacent boxes in a sealed adherent stem cell culture factory of the present application;

[0043] Figure 12 It is a structural schematic diagram of A-A section in a sealed adherent stem cell culture factory of the present application;

[0044] Figure 13 It is a sectional view structural schematic diagram of a breathable cover in a sealed adherent stem cell culture factory of the present application;

[0045] Figure 14 It is a top view structural schematic diagram of a sealing ring in a sealed adherent stem cell culture factory of the present application;

[0046] Among them, each reference sign is:

[0047] 10 - bottom box; 20 - box body, 21 - air inlet channel, 22 - air outlet channel, 23 - arc-shaped coaming, 24 - side hole; 30 - cover body, 31 - side frame, 32 - triangular connecting piece, 33 - reinforcing rib, 34 - scale, 35 - air inlet pipe, 36 - air outlet pipe, 37 - external thread; 40 - sealing mechanism, 41 - sealing column, 42 - T-shaped ejector rod, 43 - limiting ring, 44 - conical center, 45 - upper tooth column, 46 - lower tooth column, 47 - inverted L-shaped strutting rod, 48 - inverted trapezoidal buckle, 49 - return spring; 50 - gas permeable cover, 51 - internal thread, 52 - outer ring hole, 53 - inner ring hole, 54 - gas permeable film; 60 - sealing ring, 61 - sealing gasket, 62 - through hole; 70 - pressure cap. DETAILED DESCRIPTION

[0048] The application will be described in detail and specifically below through specific examples, so that the application can be better understood, but the following examples do not limit the scope of the application.

[0049] Example 1

[0050] Please refer to Figure 1 , the embodiment provides a sealed adherent stem cell culture factory, which comprises a bottom box 10, a cover body 30 and at least one box body 20 stacked between the bottom box 10 and the cover body 30; the whole sealed adherent stem cell culture factory is a layered structure composed of at least one box body 20 stacked between the bottom box 10 and the cover body 30; the bottom box 10, the box body 20 and the cover body 30 arranged in layers can be quickly buckled and connected through the sealing mechanism 40; the sealing mechanism 40 can adopt bolts or any other structure of fixing piece to ensure that the bottom box 10, the box body 20 and the cover body 30 have good sealing property through the sealing mechanism 40.

[0051] In this embodiment, please refer to Figures 4-5 , the adjacent two corners of the box body 20 are respectively provided with an air inlet channel 21 and an air outlet channel 22, and an arc-shaped coaming 23 is vertically arranged along the edges of the air inlet channel 21 and the air outlet channel 22, and a side hole 24 is formed above the arc-shaped coaming 23, which communicates the culture area of the box body 20 and the air inlet channel 21 and the air outlet channel 22.

[0052] In this embodiment, please refer to Figure 1 , the adjacent two corners of the cover body 30 are respectively provided with an air inlet pipe 35 and an air outlet pipe 36 which penetrate up and down, and the air inlet pipe 35 and the air outlet pipe 36 are respectively arranged one by one corresponding to the air inlet channel 21 and the air outlet channel 22 below.

[0053] In this embodiment, please refer to Figures 11-12As shown, the inner side walls of the bottom box 10, the box body 20 and the cover body 30 are respectively provided with a plurality of sealing mechanisms 40 corresponding in up and down direction, the bottom of the sealing mechanism 40 is provided with a movable inverted trapezoidal buckle 48, the top is provided with a buckle opening matched with the inverted trapezoidal buckle 48, the bottom box 10, the box body 20 and the cover body 30 arranged adjacent in up and down direction are connected through the buckle connection of the sealing mechanism 40, the connection has good sealing property and high stability, the yield of single batch is greatly improved, and the sealing type wall-adhering stem cell culture factory is suitable for large-scale production in a factory.

[0054] In the embodiment, the bottom box 10, the box body 20 and the cover body 30 are integrally made of transparent material, preferably, the transparent material is transparent glass fiber reinforced plastic, specifically, one or more of polypropylene (PP), polyvinyl chloride (PVC) and polystyrene (PS) and glass fiber composite material.

[0055] The sealing type wall-adhering stem cell culture factory provided in the embodiment has the working principle of wall-adhering cell culture and collection, which includes the following steps:

[0056] (1) Take the sealing type wall-adhering stem cell culture factory assembled as a whole, according to the culture specification and size of the sealing type wall-adhering stem cell culture factory, first inject the mixed solution of the prepared wall-adhering cell stock solution and culture suspension into the bottom box 10 and the box body 20 at the bottom of the cell factory through the air inlet channel 21 of the air inlet pipe 35 in a quantitative manner;

[0057] (2) Then, the cell culture factory is turned over to the front side or the back two sides and placed horizontally, the height of the two ends of the cell factory is adjusted, so that the scale line 34 on the vertical cell factory is in a horizontal state, and the mixed culture solution is equally distributed in the bottom box 10 and the box body 20 through the air inlet channel 21 and the air outlet channel 22 and the side hole 24 which are connected to each other, and the mixed culture solution is equally distributed in the bottom box 10 and the box body 20 through the air inlet channel 21 and the air outlet channel 22 and the side hole 24 which are connected to each other;

[0058] (3) After the equal distribution of the mixed culture solution, the cell factory is tilted to one side, so that the end of the cell factory with the air inlet channel 21 and the air outlet channel 22 is upward, and the mixed culture solution in the bottom box 10 and each layer of box body 20 accumulates at the bottom;

[0059] (4) Manually turn the cell factory to one side and place it horizontally, so that the mixed culture solution is evenly distributed in each layer of culture area, the equal liquid injection operation is completed, the air inlet channel 21 and the air outlet channel 22 are connected through the side hole 24, and a gas permeation channel is formed in each layer of culture area, so that the wall-adhering cells can still obtain sufficient gas permeation amount even in the full liquid culture environment, and the normal growth and culture of the cells are ensured;

[0060] (5) After the culture of the adherent cells is completed, the cover 30 is first removed by pressing the sealing structure 40 on the cover 30, then the first layer of the box body 20 is removed by pressing the sealing structure 40 on the first layer of the box body 20, the culture waste liquid in the first layer of the box body 20 is poured out, the adherent cells after culture are manually blown and collected, and the blowing and collection of the adherent cells in each layer of the box body 20 and the bottom box 10 are completed in this way.

[0061] In this embodiment, in step (5), after the box body 20 of the upper layer is removed, the cover 30 is manually assembled again, and the cell factory is sealed again by the sealing structure 40 to prevent the cultured cells in the second layer of the box body 20 from being contaminated.

[0062] In this embodiment, as a preferred scheme, in step (5), in order to quickly collect the adherent cells after culture, the whole cell factory after culture is tilted, and steps (4), (3) and (2) are repeated in reverse order to pour the culture waste liquid in each cell culture area out of the gas inlet channel 21 or the gas outlet channel 22; the cell digestion solution is injected from the gas inlet channel 21, and steps (2)-(4) are repeated to distribute the cell digestion solution in each layer of the culture area in equal amounts; after the digestion of the digestion solution is completed, the adherent cell suspension after culture is collected by pouring out of the gas inlet channel 21 or the gas outlet channel 22.

[0063] Embodiment 2

[0064] Please refer to Figures 6-11 Unlike the above-mentioned embodiment 1, this embodiment also provides a sealing mechanism 40 suitable for the box body 20 and the cover 30 of the sealed adherent stem cell culture factory. Each sealing mechanism 40 on the box body 20 and / or the cover 30 comprises a sealing column 41 arranged on the inner side wall of the box body 20 and / or the cover 30, a T-shaped ejector rod 42 vertically arranged in the accommodating cavity, and an elastic buckle member arranged in the accommodating cavity below the conical tip 44.

[0065] In this embodiment, please refer to Figure 6 As shown in the figure, the sealing column 41 on the box body 20 and / or the cover 30 is arranged on the inner side wall of the box body 20 and / or the cover 30, the sealing column 41 is provided with a hollow accommodating cavity, and the top and bottom of the sealing column 41 are respectively provided with a bayonet communicating with the accommodating cavity.

[0066] In this embodiment, please refer to Figure 6As shown, the T-shaped ejector rod 42 is vertically arranged in the accommodating chamber, the upper end of the T-shaped ejector rod 42 movably sleeved with an annular upper tooth column 45, the inner wall of the accommodating chamber is provided with an annular lower tooth column 46 matched with the upper tooth column 45, and the bottom end of the T-shaped ejector rod 42 is provided with a conical ejector tip 44, and the T-shaped ejector rod 42, the upper tooth column 45 and the lower tooth column 46 constitute a rotating telescopic mechanism. The rotating telescopic mechanism is a rotating core structure of a conventional automatic ball pen, and the structural principle is the same, which will not be described in detail here. The annular upper tooth column 45 is rotatably sleeved on the upper end of the T-shaped ejector rod 42, the annular upper tooth column 45 and the annular lower tooth column 46 are respectively provided with a ratchet structure matched with each other, and the meshing of the upper tooth column 45 and the lower tooth column 46 is incomplete meshing, only a part of the tooth tip is engaged.

[0067] The working principle of the rotating telescopic mechanism is as follows: please refer to Figures 6-11 As shown, when the upper tooth column 45 slides out of the guide groove on the lower tooth column 46 under the pressing of the T-shaped ejector rod 42, the teeth of the lower tooth column 46 will slide along the inclined surface of the engaged teeth of the upper tooth column 45 due to the action of the spring, so that the upper tooth column 45 will turn an angle under the action of the inclined surface and slide into another inclined surface without a guide groove of the tooth column 46, so that the T-shaped ejector rod 42 is clamped by the rotating telescopic mechanism after moving downward to a certain displacement; repeatedly pressing the T-shaped ejector rod 42 makes the upper tooth column 45 continue to turn an angle and slide into the next guide groove of the tooth column 46, and the T-shaped ejector rod 42 and the upper tooth column 45 thereon move upward under the action of the return spring 49.

[0068] In this embodiment, please refer to Figure 6 As shown, the elastic buckle is arranged in the accommodating chamber and below the conical ejector tip 44, the elastic buckle is composed of an inverted L-shaped straining rod 47, an inverted trapezoidal buckle 48 and a return spring 49, the inverted trapezoidal buckle 48 is located at the lower end of the accommodating chamber and is connected with the inverted L-shaped straining rod 47 at both ends, and the upper end of the inverted L-shaped straining rod 47 is arranged in the accommodating chamber through the return spring 49.

[0069] In this embodiment, please refer to Figures 10-11 As shown, the rotating telescopic mechanism composed of the T-shaped ejector rod 42, the upper tooth column 45 and the lower tooth column 46 drives the T-shaped ejector rod 42 to move upward or downward, and the T-shaped ejector rod 42 expands outward while moving downward by driving the inverted L-shaped straining rod 47 and the inverted trapezoidal buckle 48, so that the inverted trapezoidal buckle 48 is buckled in the top buckle of another sealing column 41 below.

[0070] In this embodiment, please refer to Figure 6As shown, the middle of the T-shaped ejector rod 42 is fixedly provided with a limiting ring 43, and the upper tooth column 45 is arranged on the limiting ring 43. The lower end of the conical ejector pin 44 has an inwardly recessed arc surface and is embedded between the two L-shaped spreading rods 47. The recessed arc surface can be more fitted with the inner side of the L-shaped spreading rod 47, so that the T-shaped ejector rod 42 can spread the L-shaped spreading rod 47 and the inverted trapezoidal buckle 48 on both sides to a certain angle during the process of pushing the elastic buckle downward, so that the inverted trapezoidal buckle 48 can be buckled in the top socket of the sealing column 41. Please refer to Figure 8 As shown, after the T-shaped ejector rod 42 moves upward, the pressure on the elastic buckle is removed, and the L-shaped spreading rod 47 and the inverted trapezoidal buckle 48 shrink inwardly under the action of their own elasticity to return to the original shape, and the elastic buckle moves upward and shrinks into the bottom socket of the sealing column 41 under the action of the reset spring 49.

[0071] In this embodiment, please refer to Figure 12 As shown, the inner side walls of the bottom box 10, the box body 20 and the cover body 30 are respectively provided with a plurality of sealing mechanisms 40 corresponding in up and down direction, the bottom of the sealing mechanism 40 is provided with a movable inverted trapezoidal buckle 48, and the top is provided with a socket matched with the inverted trapezoidal buckle 48. The bottom box 10, the box body 20 and the cover body 30 arranged adjacent in up and down direction are connected by the sealing mechanism 40 buckling and sealing.

[0072] Embodiment 3

[0073] Based on the above embodiments, please refer to Figures 2-3 As shown, the embodiment also provides a sealing mechanism 40 suitable for the bottom box 10 of the sealed wall-pasting stem cell culture factory. It is noted that in this embodiment, the sealing mechanism 40 of the bottom box 10 is only a sealing column 41 with a containing chamber and upper and lower sockets, and the sealing column 41 on the bottom box 10 is buckled and matched with the sealing mechanism 40 of the box body 20 above it.

[0074] In this embodiment, please refer to Figures 2-3 As shown, each of the sealing mechanisms 40 on the bottom box 10 comprises: a sealing column 41 arranged on the inner side wall of the bottom box 10, the sealing column 41 is provided with a hollow containing chamber, and the top and bottom of the sealing column 41 are respectively provided with sockets communicated with the containing chamber; and a T-shaped ejector rod 42 vertically arranged in the containing chamber, the T-shaped ejector rod 42 is abutted at the socket at the top of the containing chamber through the reset spring 49 arranged at the bottom of the containing chamber.

[0075] Embodiment 4

[0076] Based on the above embodiments, please refer to Figure 1 and Figure 13The sealed adherent stem cell culture factory of the embodiment further comprises a gas-permeable cover 50 detachably arranged on the air inlet pipe 35 and the air outlet pipe 36 respectively, and the top wall of the gas-permeable cover 50 is provided with an opening and a gas-permeable film 54 arranged inside.

[0077] In the embodiment, as shown in Figure 13 , the inner wall of the gas-permeable cover 50 is provided with an internal thread 51 matched with an external thread 37 arranged on the outer wall of the air inlet pipe 35 and the air outlet pipe 36, and the gas-permeable cover 50 is screwed on the air inlet pipe 35 and the air outlet pipe 36 through the internal thread 51 and the external thread 37.

[0078] In the embodiment, as shown in Figure 1 and Figure 13 , the top wall of the gas-permeable cover 50 is provided with an outer ring hole 52 and an inner ring hole 53 arranged concentrically, and the outer ring hole 52 and the inner ring hole 53 are respectively arranged in a ring shape. The gas-permeable film 54 is arranged at the bottom of the outer ring hole 52 and the inner ring hole 53, and the inner and outer sides of the cell culture are connected through the outer ring hole 52, the inner ring hole 53, the gas-permeable film 54, the air inlet pipe 35 and the air outlet pipe 36, forming a gas-permeable channel, so that the adherent cells are in a full-liquid culture environment and can obtain sufficient gas permeation, ensuring the normal growth and culture of the cells.

[0079] In the embodiment, as shown in Figure 1 , the side frame 31 is arranged at the position of the periphery of the cover body 30, and a plurality of triangular connecting pieces 32 are arranged at the connection between the top of the cover body 30 and the side frame 31. A plurality of reinforcing ribs 33 are arranged on the top of the cover body 30 along the length direction, improving the stability of the cover body 30.

[0080] In the embodiment, as shown in Figure 1 , the scale line 34 is arranged on the top of the cover body 30 perpendicular to the direction of the reinforcing ribs 33. When the cell culture factory is placed vertically, the scale line 34 can be used to control the amount of cell culture solution added, and the cell culture solution can be balanced in each cell culture layer through the connected culture area of the box body 20 and the side hole 24 of the air inlet channel 21 and the air outlet channel 22.

[0081] Embodiment 5

[0082] Based on the above embodiments, as shown in Figure 1 , Figure 5 and Figure 14 , the sealed adherent stem cell culture factory of the embodiment further comprises a sealing ring 60 arranged on the bottom periphery of the box body 20 and the cover body 30 respectively, and the sealing ring 60 is provided with a sealing gasket 61 at the position corresponding to the sealing mechanism 40. The sealing gasket 61 is provided with a through hole 62 corresponding to the upper and lower sockets of the sealing column 41.

[0083] Embodiment 6

[0084] Based on the above-described embodiments, please refer to Figure 1 and Figure 11 As shown in the sealing type adherent stem cell culture factory, the sealing cap 70 is matched with the T-shaped ejector pin 42 of the sealing mechanism 40. When the bottom box 10, the box body 20 and the cover body 30 are assembled, the lower end of the sealing cap 70 is manually pressed into the bayonet of the sealing column 41 to apply a downward pressure on the T-shaped ejector pin 42, so as to buckle and connect the sealing column 41 on the box body 20 and the cover body 30 with the sealing mechanism 40 below, and after the assembly is completed, the sealing cap 70 is directly embedded in the bayonet of the sealing mechanism 40 on the top of the cover body 30.

[0085] The sealing type adherent stem cell culture factory provided by the present application is a laminated structure composed of a plurality of box bodies 20 stacked between the bottom box 10 and the cover body 30, which is suitable for the culture of adherent stem cells and ensures the required culture area during the culture process. The number of layers can be adjusted by controlling the number of stacked box bodies 20, which is flexible and convenient to use. The bottom box 10, the box body 20 and the cover body 30 are arranged in layers and connected by the sealing mechanism 40, which is simple to operate, has good sealing performance and high stability, can effectively improve the yield of single batch, and is suitable for large-scale production in factories.

[0086] The specific embodiments of the present application are described in detail above, but it is only as an example, and the present application is not limited to the specific embodiments described above. Any equivalent modification and substitution of the present application made by those skilled in the art is also within the scope of the present application. Therefore, any equivalent transformation and modification made without departing from the spirit and scope of the present application should be covered within the scope of the present application.

Claims

1. A sealed adherent stem cell culture factory, characterized in that, The utility model relates to a box, which comprises a bottom box (10), a cover (30) and at least one box body (20) arranged between the bottom box (10) and the cover (30). The box body (20) is provided with an air inlet channel (21) and an air outlet channel (22) at two adjacent corners respectively, and an arc-shaped baffle (23) is vertically arranged along the edges of the air inlet channel (21) and the air outlet channel (22), and a side hole (24) is formed above the arc-shaped baffle (23) to connect the culture area of the box body (20) with the air inlet channel (21) and the air outlet channel (22); The cover (30) is provided with an air inlet pipe (35) and an air outlet pipe (36) at two adjacent corners respectively, and the air inlet pipe (35) and the air outlet pipe (36) correspond to the air inlet channel (21) and the air outlet channel (22) below respectively; The inner side walls of the bottom box (10), the box body (20) and the cover (30) are respectively provided with a plurality of sealing mechanisms (40) corresponding in the up-down direction, the bottom of the sealing mechanism (40) is provided with a movable inverted trapezoidal buckle (48), the top of the sealing mechanism (40) is provided with a buckle hole matched with the inverted trapezoidal buckle (48), and the bottom box (10), the box body (20) and the cover (30) arranged adjacent in the up-down direction are connected through the buckle connection of the sealing mechanism (40); Each sealing mechanism (40) on the box body (20) and / or the cover (30) comprises: a sealing column (41) arranged on the inner side wall of the box body (20) and / or the cover (30), a hollow accommodating cavity is arranged in the sealing column (41), and the top and the bottom of the sealing column (41) are respectively provided with a buckle hole communicated with the accommodating cavity; a T-shaped ejector rod (42) vertically arranged in the accommodating cavity, an annular upper tooth column (45) is movably arranged on the upper end of the T-shaped ejector rod (42), an annular lower tooth column (46) is arranged on the inner wall of the accommodating cavity and matched with the upper tooth column (45), a conical tip (44) is arranged on the bottom end of the T-shaped ejector rod (42), and the T-shaped ejector rod (42), the upper tooth column (45) and the lower tooth column (46) constitute a rotary telescopic mechanism; and an elastic buckle piece arranged in the accommodating cavity and below the conical tip (44), the elastic buckle piece is composed of an inverted L-shaped straining rod (47), an inverted trapezoidal buckle (48) and a return spring (49), the inverted trapezoidal buckle (48) is arranged at the lower end buckle hole of the accommodating cavity, and the two ends of the inverted trapezoidal buckle (48) are connected with the inverted L-shaped straining rod (47) respectively, and the upper end of the inverted L-shaped straining rod (47) is arranged in the accommodating cavity through the return spring (49). The rotating telescopic mechanism composed of the T-shaped ejector rod (42), the upper tooth column (45) and the lower tooth column (46) drives the T-shaped ejector rod (42) to move up or down, and the T-shaped ejector rod (42) is expanded outward by the tapered ejector tip (44) at the lower end thereof while pushing the inverted L-shaped spreader rod (47) and the inverted trapezoidal buckle (48) to move down, so that the inverted trapezoidal buckle (48) is buckled in the top buckle of another sealing column (41) below it; The lower end of the tapered ejector tip (44) has an arc surface recessed inward, and is embedded between the two L-shaped spreader rods (47); The T-shaped ejector rod (42) is fixedly provided with a limiting ring (43) in the middle, and the upper tooth column (45) is arranged on the limiting ring (43); It also includes a breathable cover (50) which is detachably arranged on the air inlet pipe (35) and the air outlet pipe (36), respectively, and the top wall of the breathable cover (50) is provided with an opening and a breathable film (54) arranged inside. Each of the sealing mechanisms (40) on the bottom box (10) comprises: A sealing column (41) arranged on the inner side wall of the bottom box (10), the sealing column (41) is provided with a hollow containing chamber, and the top and bottom of the sealing column (41) are respectively provided with a buckle which communicates with the containing chamber; and A T-shaped ejector rod (42) vertically arranged in the containing chamber, the T-shaped ejector rod (42) is abutted at the buckle at the top of the containing chamber by the reset spring (49) arranged at the bottom of the containing chamber.

2. The sealed adherent stem cell culture factory of claim 1, wherein, The side frame (31) is arranged at the position of the four peripheral edges of the cover body (30), and a plurality of triangular connectors (32) are arranged at the connection between the top of the cover body (30) and the side frame (31).

3. The sealed adherent stem cell culture factory of claim 1, wherein, The top of the cover body (30) is provided with a plurality of parallel arranged reinforcing ribs (33) along the length direction thereof, and the top is provided with a scale line (34) perpendicular to the direction of the reinforcing ribs (33).

4. The sealed adherent stem cell culture factory of claim 1, wherein, It also includes a sealing ring (60) arranged at the bottom periphery of the box body (20) and the cover body (30), respectively.

5. The sealed adherent stem cell factory of claim 1, wherein, It also includes a pressure cap (70) cooperating with the T-shaped ejector rod (42) of the sealing mechanism (40).

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