CN117187063A

CN117187063A - Tangential flow bioreactor

Info

Publication number: CN117187063A
Application number: CN202311359369.7A
Authority: CN
Inventors: 高毅; 李阳; 黎煦程
Original assignee: Guangdong Qianhui Biotechnology Co ltd
Current assignee: Guangdong Qianhui Biotechnology Co ltd
Priority date: 2023-10-18
Filing date: 2023-10-18
Publication date: 2023-12-08
Anticipated expiration: 2043-10-18

Abstract

The application discloses a tangential flow bioreactor, comprising: the tank body and the tank cover are cylindrical and are sealed and fixed when being covered; the net basket is in a cylinder shape and is fixed in the tank body, and a gap is kept between the net basket and the tank bottom and the tank wall; the sampling turntable is movably covered on the top of the basket, a handle penetrating out of the tank cover is arranged in the center of the sampling turntable, and a movable hanging rod for fixing a sample wafer is arranged on the sampling turntable body along the radial direction and the circumferential direction; the bubble generator is arranged at the bottom of the tank. The tangential flow bioreactor mainly utilizes the bubble generator arranged at the bottom of the tank to provide fluid power, bubbles float upwards from the bottom of the tank and synchronously drive fluid to flow upwards, thus not only realizing tangential flow relative to microcarriers, but also having better oxygen dissolving effect of the culture medium.

Description

Tangential flow bioreactor

Technical Field

The application relates to the field of cell culture, in particular to a tangential flow bioreactor.

Background

The bioreactor, especially for large-scale culture of mammalian cells in vitro, is difficult to produce in higher orders of magnitude due to the adherence requirement and passage limitation of the mammalian cells, and the microcarrier is used as a culture carrier for adherence of the mammalian cells, so that the cell density in the bioreactor can be effectively improved, but the problem of nonuniform environment in the reactor can not be well overcome in the prior art, and the nonuniform environment in the reactor is mainly reflected in nonuniform dissolved oxygen and nonuniform update of a liquid culture medium, thereby causing inconsistent cell states in the reactor and affecting the overall quality of cells and the yield and quality of metabolites.

Disclosure of Invention

The object of the present application is to at least partially overcome the disadvantages of the prior art and to provide a tangential flow bioreactor based on microcarrier use to better achieve homogeneity within the reactor.

In order to achieve the technical purpose, the application adopts the following technical scheme:

a tangential flow bioreactor, comprising:

the tank body and the tank cover are cylindrical and are sealed and fixed when being covered;

the net basket is in a cylinder shape and is fixed in the tank body, and a gap is kept between the net basket and the tank bottom and the tank wall;

the sampling turntable is movably covered on the top of the basket, a handle penetrating out of the tank cover is arranged in the center of the sampling turntable, and a movable hanging rod for fixing a sample wafer is arranged on the sampling turntable body along the radial direction and the circumferential direction;

the bubble generator is arranged at the bottom of the tank.

Preferably, the device further comprises a sampling window which is radially arranged on the tank cover and is sealed and fixed with the tank cover when the sampling window is in a closed state.

Optionally, the sampling window adopts a flip-type structure or a sliding-type structure.

Preferably, the sampling window is at least sized to accommodate the removal of a radially disposed row of said movable hanging bars from said sampling carousel.

Further, the diameter of the air bubbles generated by the air bubble generator is less than 1mm, and the density of the holes of the basket is adapted to block the air bubbles with the diameter of more than 1 mm.

Optionally, the basket is fixed to the tank wall by a hanging assembly or placed on the tank bottom by a support leg.

Further preferably, a plurality of positioning strips are annularly distributed at the bottom of the basket along the center.

Optionally, a basket attachment is also included for hanging in the basket for fixing the culture carrier.

Preferably, the handle is configured in a circular shape and the handle surface and can lid surface are provided with circumferential locating marks.

Further, the handle is connected with the sampling turntable body through a vertical connecting rod, and the vertical connecting rod and the tank cover are provided with a detachable sealing assembly.

Alternatively, the sealing assembly is a flange type sealing assembly or a hoop type sealing assembly.

Preferably, the sampling turntable body comprises a plurality of radial connecting rods, and positioning grooves for hanging the movable hanging rods are formed in the connecting rods; the middle upper part of the movable hanging rod is provided with a hook which is opened downwards, and the top of the movable hanging rod is provided with a hanging ring.

Preferably, each connecting rod is provided with a positioning mark corresponding to each other on the handle; and each hanging ring of the movable hanging rod is provided with a positioning mark.

Further preferably, each movable hanging rod is provided with a plurality of clips for fixing the sample wafer along the axial direction of the movable hanging rod; the clip is provided with a positioning mark.

Still further, the tank also comprises a bubble generator arranged on the wall of the tank.

Compared with the prior art, the application has the following advantages:

(1) The tangential flow bioreactor mainly utilizes the bubble generator arranged at the bottom of the tank to provide fluid power, bubbles float upwards from the bottom of the tank and synchronously drive fluid to flow upwards, thus not only realizing tangential flow relative to microcarriers, but also having better oxygen dissolving effect of the culture medium.

(2) The tangential flow bioreactor disclosed by the application has the advantages that the basket can be used as a support of a culture carrier, bubbles with the size above can be blocked, the damage of large bubbles to cells attached to the surface of the culture carrier is avoided, and the protection to the cells in culture is realized.

(3) When the tangential flow bioreactor adopts the microcarrier coiled material as a culture carrier, the coiled material can stand in the basket by itself, and when the laminar or non-stand culture carrier is adopted, the coiled material can be maintained in the basket by the aid of various basket accessories, so that the tangential flow bioreactor can be suitable for various culture carriers, and the applicability of the tangential flow bioreactor is improved.

(4) The tangential flow bioreactor adopts a cylindrical design, and utilizes the central symmetry characteristic of a cylinder to arrange a sampling rotary table and a corresponding sampling window, thereby solving various problems of comprehensiveness, convenience and pollution resistance of sampling.

Drawings

FIG. 1 is a schematic view of the longitudinal structure of a tangential flow bioreactor according to the present application.

Fig. 2 is a schematic view of the transverse structure of the tangential flow bioreactor of the present application, in which a part of the structure of the tank cover, a part of the structure of the basket and a part of the structure of the tank bottom are shown.

Fig. 3 is an enlarged schematic view of the area a in fig. 2.

Fig. 4 is an enlarged schematic view of region B in fig. 1.

Detailed Description

The application is described in further detail below with reference to the drawings and detailed description.

Referring to fig. 1 and 2, the tangential flow bioreactor of the present application comprises a cylindrical tank 1 and a tank cover 2, a basket 3 fixed in the tank 1, a sampling turntable 4 covered on the top 31 of the basket, and a bubble generator.

Specifically, the tank 1 and the tank cover 2 are sealed when in use, so as to provide a stable internal environment isolated from the outside for the cells in the tank, and the tank 1 and the tank cover 2 need to be provided with necessary substance exchange channels, such as an inlet and an outlet of a culture solution, and also need to be provided with necessary detection channels, such as a pH detection channel, a dissolved oxygen detection channel, a sugar content detection channel, and the like, respectively. Further, the tank 1 has an accommodation space defined by the tank bottom 11 and the tank wall 12, in which the basket 3 is fixed in the present application, the basket 3 occupies most of the accommodation space for use as a cell culture site, and preferably the basket 3 needs to be kept in a gap with the tank bottom 11 and the tank wall 12. One possibility is to provide a hooking component 32 between the upper edge of the basket 3 and the tank wall 12 for fixation, for example a hooking element with the basket 3 provided with a downward return hook, and correspondingly the tank wall 12 provided with a support with an upward return hook, in which the hooking element can be placed from above downwards, so that the bottom of the basket 3 is kept at a predetermined distance from the tank bottom 11, and the hooking component 32 also has a predetermined width, so that the side wall of the basket 3 is kept at a predetermined distance from the tank wall 12. Another possibility is that the bottom of the basket 3 is provided with a plurality of supporting feet, the basket 3 is fixed by the bottom of the basket 3 and the tank bottom 11, and the basket 3 fixed by the supporting feet and the tank bottom 11 and the tank wall 12 respectively keep preset gaps.

Further, the basket 3 of the present application is used for loading microcarriers for culturing cells having an adherent characteristic, and is advantageously applied to a sheet-like carrier based on polyester fiber, which has an internal structure and a surface structure interlaced into a net, and which can be manufactured in a multi-layer structure to further expand the surface area for adherence, and commercially available sheet-like carriers are classified into a sheet-like carrier of a coil shape, a sheet-like carrier of a short strip shape, and a sheet-like carrier of a polygonal shape according to the size based on the manufacturing process of the sheet-like carrier.

When the basket 3 of the present application is used for loading the coiled sheet-like carriers, the coiled sheet-like carriers are required to be vertically placed in the basket 3 as a whole, so that the coiled sheet-like carriers are axially parallel to the basket 3, and the coiled sheet-like carriers should maintain a certain gap between layers so as to facilitate the passage of the culture medium through each sheet-like carrier, preferably, a plurality of positioning strips 33 are annularly distributed along the center at the bottom of the basket 3 of the present application for assisting in defining the interlayer gap of the sheet-like carriers. If the web-like sheet-like carrier is difficult to erect in a vertical state with its own stiffness, some supports, such as rod-like supports, clip-like supports, may be provided in the upper middle portion of the basket 3. In another implementation, a portion of the positioning strips 33 may extend downwardly from your bottom of the basket 3 to form the support feet described above.

When the basket 3 according to the application is used for loading short-strip-shaped sheet carriers and polygonal sheet carriers, the sheet carriers are preferably kept in a vertical state, i.e. perpendicular to the bottom of the basket 3/tank bottom 11, since discrete sheet carriers cannot be erected with their own stiffness support, it is necessary to provide adapted basket 3 attachments for loading sheet carriers, preferably various basket 3 attachments can be fixed in the basket 3 by means of hanging. The accessories of the basket 3 can be configured into hanging rods, hanging hooks 441, hanging clamps and the like with hanging functions according to different functions, hanging bags and the like with wrapping or semi-wrapping functions, and the specific forms of the accessories of the basket 3 can be improved in adaptability.

The basket 3 of the present application is also suitable for spherical microcarriers, but the hole density of the basket 3 needs to be adapted to block spherical microcarriers from escaping outside the net.

The application adopts a bubble generator to synchronously provide gas (oxygen and carbon dioxide) and hydrodynamic force required by cells. The bubble generator is roughly classified into a centimeter bubble (1 mm or more), a millimeter bubble (100 μm to 1 mm), a micrometer bubble (1 μm to 100 μm), a micrometer bubble (tens nm to 1 μm) and a nanobubble (tens nm or less) according to the bubble diameter generated. In practical applications, millimeter bubbles, microbubbles, and micro-nanobubbles are suitable for application to living organisms, but these three bubbles have distinct physical forms: millimeter bubbles (containing sub-millimeter bubbles) are visible, slowly rise in the liquid, and collapse when reaching the water surface; the micro-bubbles are visible visually, are in a white turbid state, can rise very slowly in the liquid, and disappear in the water along with the time extension; the ultra-fine bubbles are invisible and colorless and transparent, and can remain in water for several weeks to months, and do Brownian movement in water. By combining the requirements of the tangential flow bioreactor, the oxygen dissolving effect of the culture medium can be improved, the culture medium can be driven to move upwards, preferably three different-size bubbles are adopted, namely three different-size bubbles can be correspondingly produced by adopting three different bubble generators, or one bubble generator capable of producing three different-size bubbles can be adopted, and the method is not limited in practical application; secondly, if only one size of bubbles is considered, it is preferable to use a bubble generator for producing microbubbles.

The bubble generator suitable for the application comprises a bubble aeration head 5, a host (not shown) and a gas tank (not shown), wherein the host and the gas tank are arranged outside the tank body 1, and the working principle of the bubble generator can be as follows:

spinning liquid flow-liquid is pumped into the cylindrical generator body from its side in tangential direction to produce high speed rotational flow inside. By using the pressure drop generated by the liquid rotation movement, the self-sucking gas in the small hole at the lower end surface is crushed under the action of the shearing force in the small hole at the center of the upper end surface to form micro bubbles.

Pressurized dissolution-pressurizing a gas-liquid mixture with a pump to dissolve the gas component in the liquid to supersaturation. Undissolved gas bubbles are separated, and only supersaturated liquid enters normal pressure liquid through a pressure reducing valve to precipitate micro-bubbles.

Micropore-gas injected into a liquid is sheared by a high-speed liquid flow along the microporous membrane face to microbubbles.

The bubble aeration head 5 of the bubble generator of the present application is preferably disposed outside the tank bottom 11, the basket 3, i.e., in the gap between the basket 3 and the tank bottom 11, more preferably at a position where the positioning bar 33 of the basket 3 is not blocked (as shown in fig. 2). The number of the bubble aeration heads 5 and the amount of bubbles of the culture medium injected into the bubble generator are required to be adjusted according to the cell experiment, and if a plurality of bubble aeration heads 5 are arranged at the tank bottom 11, the plurality of bubble aeration heads 5 are communicated by using a gas pipe. Further, the bubble aeration heads 5 may be further disposed outside the tank wall 12 and the basket 3, that is, in the gap between the basket 3 and the tank wall 12, where the number of the bubble aeration heads 5 on the tank wall 12 and the total amount of bubbles need to be adjusted by matching with the setting parameters of the bubble aeration heads 5 of the tank bottom 11 through a cell experiment, and the bubble aeration heads 5 of the tank wall 12 may be communicated with the bubble aeration heads 5 of the tank bottom 11 through a gas pipe.

The bubble generator has advantages for cell culture, but the bubbles are easy to generate mechanical damage to animal cells without cell walls, and large bubbles are not expected to be generated by the application, so that the bubbles with the pore density of the basket 3 being more than millimeter size are blocked by the adaptation of the pore density, on one hand, the large bubbles can be blocked, and on the other hand, the bubbles of the bubble generator are prevented from directly striking the opposite cells.

In addition, as will be appreciated by those skilled in the art, the oxygen requirement of animal cells is not pure oxygen, so the gas used in the present application should comprise two kinds of oxygen and carbon dioxide, which are proportioned and then injected into the culture medium by forming microbubbles through a bubble generator.

The sampling turntable 4 is movably covered on the top 31 of the basket 3, and the sampling turntable 4 can be horizontally rotated on the top 31 of the basket 3 by taking the center of the sampling turntable as an axis, so that any piece 6 fixed on the sampling turntable 4 can be horizontally rotated back to the position below the sampling window 21 of the tank cover 2 and can be taken out. A possible implementation way is that the top 31 of the basket 3 is provided with a chute for accommodating the edge of the sampling turntable 4 in a whole circle so as to facilitate the stable and flat rotation of the sampling turntable 4; in another implementation manner, the edge of the sampling turntable 4 is provided with a buckling structure of a downward half bag so as to be buckled on the smooth top 31 of the basket 3 for stable and flat rotation. The sampling turret 4 according to the application is preferably provided with several links based on the central radiation, all of which constitute the body 41 of the sampling turret 4, the radiation center being provided with a handle 42 penetrating out of the can lid 2 by means of a vertical link 43, whereby the sampling turret 4 can be translated outside the can lid 2 by rotating the handle 42. Further, a movable hanging rod 44 is hung on the connecting rod along the radial direction, the movable hanging rod 44 penetrates into the bottom of the basket 3, and a plurality of clips 443 for clamping the sample wafer 6 are arranged.

Specifically, referring to fig. 3 and 4, the sampling turntable 4 is provided with 6 to 8 links, or a greater number of links for increasing sampling density, and the links are uniformly distributed on the circle defined by the basket 3. A hanging structure is arranged between the connecting rod and the movable hanging rod 44, one possible implementation mode is that a positioning slot 411 is formed in the connecting rod, a hook 441 which is downwards opened is arranged on the movable hanging rod 44, the movable hanging rod 44 is fixed in the positioning slot 411 through the hook 441 after moving into the positioning slot 411, and in order to improve flexibility, the positioning slot 411 has a certain length, one implementation mode is that the positioning slot 411 is a waist-shaped hole, one side of the waist-shaped hole is communicated with the longitudinal edge of the connecting rod to form an opening, the movable connecting rod enters the positioning slot 411 through the opening, and the hook 441 is fixed at any position on two sides of the waist-shaped hole; further, to enhance the positioning effect, the edges of the waist-shaped hole may be optimized to be wavy lines or continuous U-shaped lines, so as to avoid the deviation of the hook 441 in the positioning slot 411. Preferably, a plurality of positioning slots 411 are radially arranged on one connecting rod, so that a plurality of movable hanging rods 44 can be hung on one connecting rod to represent the growing environment of the cells with different interlayer positions.

Referring to fig. 1 and 4, the movable hanging rod 44 is preferably a flat rod, the top of the movable hanging rod is provided with a hanging ring 442, a hanging hook 441 for hanging the body 41 of the sampling turntable is arranged near the top of the movable hanging rod, and a plurality of clips 443 are uniformly arranged below the hanging hook 441 on the rod body at intervals. The clip 443 can be matched with a through hole, a blind hole or a sliding groove extending in the longitudinal direction formed on the body of the movable hanging rod 44, and the clip 443 can be repeatedly detached, so that the convenience of use is improved, and the structure of being convenient for one-hand assembly and disassembly can be provided. When in use, a microcarrier sample wafer 6 with proper size is cut, and the microcarrier sample wafer 6 is fixed at different positions of the movable hanging rod 44 by using the clip 443, so as to represent the cell growth environments with different depths. In a simple design, the clip 443 can be replaced with a disposable gun head, which has the advantage of reducing the replacement cost of the special clip 443, and the disposable gun head is convenient for sterilization and replacement.

Thus, the sampling turntable 4 according to the present application can be provided with a plurality of different sampling sheets 6 in the accommodation space of the basket 3 according to the distribution position of the clips 443, and each sampling sheet 6 can be positioned in three dimensions of circumferential, radial and depth.

The tank cover 2 is used for keeping the internal and external isolated state of the tank body 1, preferably the tank cover 2 should keep sealing when being covered with the tank body 1, firstly the position where the tank cover 2 contacts with the tank body 1 is sealed by a conventional way, secondly the sampling turntable 4 is provided with the handle 42 penetrating out of the tank cover 2, then a gap exists between the tank cover 2 and the vertical connecting rod 43 connected with the handle 42, the solution of the application is that a detachable sealing component is arranged between the tank cover 2 and the vertical connecting rod 43, so that the sealing component is detached or loosened when the handle 42 needs to be rotated, alternatively, the sealing component adopts a flange type sealing component, specifically, a flange structure is arranged at the central opening of the tank cover 2, an adaptive flange structure is arranged at the periphery of the vertical connecting rod 43, when the sealing is needed, the two flange structures are tightly connected, and a sealing layer is needed between the two flange structures. In another implementation manner, the sealing assembly is a hoop type sealing assembly, and the hoop type sealing assembly is generally composed of two semicircular half hoops, so that a sealing layer filled between the tank cover 2 and the vertical connecting rod 43 can be pressurized when the two half hoops are folded and connected, and a sealing effect is achieved. The flange type sealing assembly and the hoop type sealing assembly are in a disassembly state, so that the horizontal rotation of the sampling turntable 4 is not influenced, namely, when sampling is required to be carried out through the horizontal rotation sampling turntable 4, the sealing assembly is required to be disassembled, and when the sealing assembly is assembled, the sampling turntable 4 and the tank cover 2 are fixed, and the sampling turntable 4 cannot horizontally rotate relative to the tank cover 2.

Preferably, the can lid 2 is provided with a sampling window 21, the sampling window 21 is radially opened on the can lid 2, and the sampling window 21 is sealed and fixed with the can lid 2 when in a closed state, for maintaining the isolation function of the can lid 2. Alternatively, the sampling window 21 may be in a flip-type structure or a slide-type structure (as shown in fig. 1 and 2), and the opening size of the sampling window may be at least matched with the requirement that a row of movable hanging rods 44 distributed on the sampling turntable 4 along the radial direction can be taken out, that is, the opening of the sampling window 21 covers at least the movable hanging rods 44 on the connecting rod of one sampling turntable 4. In the application, if the sampling window 21 is not arranged on the tank cover 2, the tank cover 2 needs to be opened entirely when the sample wafer 6 is taken out, thus being inconvenient to operate and increasing the risk of pollution of cells; if a plurality of sampling windows 21 are arranged, the cleaning difficulty and the sterilization difficulty of the tank cover 2 are increased, the risk of pollution to cells is increased, the space for arranging other channels on the tank cover 2 is also occupied, and the plurality of sampling windows 21 are not beneficial to the sampling turntable 4 of the application. The tank cover 2 is only provided with one sampling window 21, any sample piece 6 is required to be taken out, the movable hanging rod 44 where the target sample piece 6 is located is only required to be rotated to be horizontally rotated below the sampling window 21 by rotating the handle 42, the movable hanging rod 44 where the sample piece 6 is located can be taken out through the sampling window 21, the target sample piece 6 is taken off from the movable hanging rod 44, or the sample piece 6 is replaced, the movable hanging rod 44 and the rest sample pieces 6 can be continuously sampled as required, or the movable hanging rod 44 is hung on the original connecting rod again, and after the movable hanging rod 44 is hung on the original connecting rod, the sampling turntable 4 is required to be horizontally rotated and reset, so that the position of the rest sample piece 6 is prevented from being changed.

Since each sample 6 can be positioned in three dimensions of circumferential, radial and depth, and the sample 6 needs to be transferred to the sampling window 21 for resetting when sampling, the application sets a positioning mark 7 system in the reactor, comprising:

the connecting rod-handle-can cover mark group is mainly arranged on the handle 42 and the can cover 2 in one-to-one correspondence with the positioning marks 7, and the marks on the handle 42 represent the positions of the connecting rods because the connecting rods and the handle 42 cannot rotate relatively in a flat mode, so that the rotating handle 42 inevitably drives the connecting rods to rotate synchronously. In one possible implementation, referring to fig. 2, the round surface of the can lid 2 is divided into eight parts, each part of lid edge is provided with a mark, adjacent marks are different, correspondingly, the embodiment shown in fig. 2 is provided with eight connecting rods, correspondingly, the round surface of the handle 42 is divided into eight parts, each part of handle 42 edge is provided with a mark, adjacent marks are different, but the can lid marks and the handle marks in the same sector are the same, so as to form a mark pair; the sampling window 21 is arranged on the radial line where one of the marker pairs is located;

a set of eye marks, positioning marks 7 disposed on the eye 442 of the movable boom 44; at least one movable hanging rod 44 is hung on the same connecting rod, and different movable hanging rods 44 represent interlayer positions with different distances relative to the circle center;

a clip mark group, a positioning mark 7 arranged on the clip 443 of the movable hanging bar 44; a plurality of clips 443 are provided on the same movable hanger bar 44, and different clips 443 represent positions of different depths relative to the surface of the medium.

Implementations of the positioning mark 7 include, but are not limited to, one, two or a combination of two or more of the following: color, number, letter, chinese character, foreign language, pattern, shape and pattern. The size of the positioning mark 7 is preferably clearly visible, and the surface on which the positioning mark 7 is provided may be subjected to a finishing treatment to increase the visibility and the recognition.

When the reactor is used for static culture, each mark pair in the mark group of the connecting rod-handle 42-tank cover 2 is matched with each other, and the mark pair at the sampling window 21 can be used as a reset mark; during sampling, the position of the target sample wafer 6 is determined, the center of the circle of the body 41 of the sampling turntable can be used as an origin, the coordinates of the sample wafer 6A are recorded as A (a 'hanging ring mark', 'handle-can cover mark pair', 'clip mark'), and the usage of a cylindrical coordinate system can be referred to. Sampling is performed in a sterile environment. To obtain the target specimen 6, the handle mark in the target mark pair is firstly turned to the sampling window 21 according to the handle-can lid mark pair; the sampling window 21 may then be opened after the handle 42 is turned flat into place; the second step is to hook the movable hanging rod 44 at the corresponding position from the tank according to the hanging ring mark; thirdly, removing the clips 443 at the corresponding positions according to the clip marks to obtain a target sample 6; fourth, the movable hanging rod 44 is reset, if the movable hanging rod 44 is not needed to be put into the tank body 1 again, the sampling rotary table 4 can be reset directly, and the sampling window 21 should be covered and sealed before the handle 42 is turned horizontally.

According to the requirement of aseptic operation, the sampling window 21 should be covered when the handle 42 is turned, that is, if sampling of a plurality of samples 6 at different positions is to be performed, after the current target sample 6 is removed, the sampling window 21 should be covered first, then the handle 42 is turned to transfer the next target sample 6 under the sampling window 21, and the sampling window 21 is opened again before the next movable hanging rod 44 is taken out. When further improving the requirement of aseptic operation, the sampling window 21 should be covered in time after the movable hanging rod 44 is taken out, and the sampling window 21 needs to be re-opened before the movable hanging rod 44 is placed in the tank body 1. Thus, the movable part of the sampling window 21 should be adapted to be frequently opened and closed, and the assembly for sealing should be simple in structure but excellent in sealing property.

In summary, the tangential flow bioreactor of the present application comprises: the tank body and the tank cover are cylindrical and are sealed and fixed when being covered; the net basket is in a cylinder shape and is fixed in the tank body, and a gap is kept between the net basket and the tank bottom and the tank wall; the sampling turntable is movably covered on the top of the basket, a handle penetrating out of the tank cover is arranged in the center of the sampling turntable, and a movable hanging rod for fixing a sample wafer is arranged on the sampling turntable body along the radial direction and the circumferential direction; the bubble generator is arranged at the bottom of the tank. The tangential flow bioreactor mainly utilizes the bubble generator arranged at the bottom of the tank to provide fluid power, bubbles float upwards from the bottom of the tank and synchronously drive fluid to flow upwards, thus not only realizing tangential flow relative to microcarriers, but also having better oxygen dissolving effect of the culture medium.

The above embodiments are preferred embodiments of the present application, but are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principles of the present application should be made therein and are intended to be equivalent substitutions within the scope of the present application.

Claims

1. A tangential flow bioreactor, comprising:

the bubble generator is arranged at the bottom of the tank.

2. The tangential flow bioreactor of claim 1, further comprising a sampling window radially disposed on the lid, the sampling window being sealingly secured to the lid when in a closed position.

3. The tangential flow bioreactor of claim 2, wherein the sampling window is in a flip-type structure or a slide-type structure.

4. A tangential flow bioreactor as set forth in claim 3, wherein the sampling window is at least sized to accommodate removal of a radially disposed array of said movable hanging bars from said sampling rotor.

5. The tangential flow bioreactor of claim 1, wherein the bubble generator produces bubbles having a diameter of less than 1mm, and wherein the basket has a density of holes adapted to block bubbles having a diameter of greater than 1 mm.

6. The tangential flow bioreactor of claim 1, wherein the basket is secured to the tank wall by a hanging assembly or is placed on the tank bottom by a support foot.

7. The tangential flow bioreactor of claim 1, wherein the basket bottom is annularly distributed with a plurality of locator bars along the center.

8. The tangential flow bioreactor of claim 1, further comprising a basket attachment for hanging within the basket for securing the culture carrier.

9. The tangential flow bioreactor of claim 1, wherein the handle is configured as a circle and the handle surface and the lid surface are provided with circumferential locating marks.

10. The tangential flow bioreactor of claim 1, wherein the handle is coupled to the sampling carousel body by a vertical link, the vertical link providing a removable seal assembly with the tank cap.

11. The tangential flow bioreactor of claim 10, wherein the seal is a flange seal or a hoop seal.

12. The tangential flow bioreactor of claim 1, wherein the sampling turntable body comprises a plurality of radial connecting rods, and positioning grooves for hanging the movable hanging rods are formed in the connecting rods; the middle upper part of the movable hanging rod is provided with a hook which is opened downwards, and the top of the movable hanging rod is provided with a hanging ring.

13. The tangential flow bioreactor of claim 12, wherein each of the links has a one-to-one locating mark on the handle; and each hanging ring of the movable hanging rod is provided with a positioning mark.

14. The tangential flow bioreactor as set forth in claim 1 or 12, wherein each of the movable hanger bars is provided with a plurality of clips for fixing the sample along an axial direction thereof; the clip is provided with a positioning mark.

15. The tangential flow bioreactor of claim 1, further comprising a bubble generator disposed on the tank wall.