Point vibration table suitable for culturing adherent cells

Abstract

The invention belongs to the technical field of cell culture, and relates to a point vibration table suitable for culturing adherent cells, which comprises a point vibration table top provided with a point vibration through hole; a groove matched with the point vibration through hole is formed in the side surface of the elastic body; the elastic body is installed in the point vibration through hole in an embedded mode, and the groove accommodates the periphery of the point vibration through hole. The eccentric shaft comprises a main shaft and an output shaft, and the output shaft and the main shaft are not coaxial; one end of the connecting rod mechanism is mounted on the output shaft through a bearing, and a point vibration rod is mounted at the other end; when the main shaft rotates, the point vibration rod only performs linear motion perpendicular to the lower surface of the elastic body and performs point vibration on the elastic body; and the driving mechanism drives the main shaft of the eccentric shaft to rotate. According to the application, the cells are dispersed by beating the surface of the cell factory in the vertical direction, so that the viability of the cells is ensured, and a relatively good dispersion effect is achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of cell culture, relates to the realization of digestion and passage in a cell scale culture process, and specifically relates to a point vibration platform suitable for culturing adherent cells. Background Art

[0002] When adherent cells are cultured, they adhere to the surface of the cell factory to achieve continuous proliferation. When the iron-walled cells proliferate to the point where they cover the entire surface of the cell factory, the cells in the cell factory can be digested and collected.

[0003] After the cells are collected, they are centrifuged and the supernatant is removed. Many cells are still in a state of clumps or pieces, and it is necessary to separate the cell clumps by shaking.

[0004] Currently, existing mechanical cell dispersion processes are based on the principle of generating centrifugal force in the horizontal direction to promote cell dispersion; or based on eddy current dispersion to promote cell dispersion, such as JP7064191 B2 cell dispersion method and cell disperser, which adopt eddy current dispersion technology.

[0005] However, due to the particularity of adherent cell culture, the above two methods cannot assist the detachment of adherent cells cultured in cell factories, nor can they guarantee the viability and aggregation rate of cells after detachment. Summary of the invention

[0006] The present application provides a point vibration table suitable for culturing adherent cells, which achieves cell dispersion by vertically tapping the surface of the cell factory, thereby ensuring the cell viability and achieving a better dispersion effect.

[0007] In order to achieve the above technical purpose, the technical solution adopted in the present application is a point vibration platform suitable for culturing adherent cells, comprising

[0008] The point-vibration table is provided with a point-vibration through hole;

[0009] An elastic body, a groove matching the point-vibration through hole is provided on its side; the elastic body is embedded in the point-vibration through hole, and the groove accommodates the periphery of the point-vibration through hole;

[0010] An eccentric shaft comprises a main shaft and an output shaft, wherein the output shaft is not coaxial with the main shaft;

[0011] A connecting rod mechanism, one end of which is mounted on the output shaft through a bearing, and the other end of which is mounted with a point-shock rod; the point-shock rod is located on the lower surface of the elastic body; when the main shaft rotates, the point-shock rod only moves linearly perpendicular to the lower surface of the elastic body and performs point-shock on the elastic body;

[0012] A driving mechanism drives the main shaft of the eccentric shaft to rotate.

[0013] As an improved technical solution of the present application, the point vibration table is provided with two point vibration through holes; there are two elastomers, each of which is embedded and installed in a corresponding point vibration through hole; the eccentric shaft includes a main shaft and output shafts located at both ends of the main shaft; the axes of the two output shafts are respectively parallel to the axis of the main shaft, and are axially symmetrical with respect to the main shaft; each output shaft is connected to one of the connecting rod mechanisms; each of the connecting rod mechanisms performs point vibration on one of the elastomers.

[0014] As an improved technical solution of the present application, the point shock table is provided with two groups of point shock through holes; each group of point shock through holes is composed of a plurality of point shock through holes; there are two groups of elastomers, and the number of elastomers in each group corresponds to the number of point shock through holes in each group; the eccentric shaft includes a main shaft and output shafts located at both ends of the main shaft; the axes of the two output shafts are respectively parallel to the axis of the main shaft, and are axially symmetrical relative to the main shaft; each output shaft is connected to a connecting rod mechanism; the point shock rod in each connecting rod mechanism contains a plurality of point shock points, and the number of point shock points in each connecting rod mechanism corresponds to a group of elastomers; the point shock points complete the point shock of the elastomer.

[0015] As an improved technical solution of the present application, the point shock rod includes

[0016] There are multiple point shock points, which correspond one by one to a group of elastic bodies;

[0017] A connecting rod connects all the point-vibration points so that all the point-vibration points move linearly at the same time;

[0018] A point shock rod support body, one end of which is driven by the output shaft through the connecting rod mechanism, and the other end of which supports the connecting rod.

[0019] As an improved technical solution of the present application, the connecting rod mechanism includes

[0020] The first connecting member is in an L-shaped structure, and a first mounting hole and a second mounting hole are respectively provided at two ends of the L-shaped structure;

[0021] A second connecting member, one end of which is provided with a first bearing mounting hole, and the other end of which is provided with a second bearing mounting hole; a bearing is installed in the first bearing mounting hole, and the output shaft of the eccentric shaft is installed in the first bearing mounting hole; another bearing is installed in the second bearing mounting hole, and the first mounting hole and the second bearing mounting hole are connected by a connecting shaft;

[0022] One end of the point shock rod is fixed on the second mounting hole and supported by the second mounting hole; the other end is a point shock point for vibrating the elastic body.

[0023] As an improved technical solution of the present application, the elastomer is made of silicone.

[0024] As an improved technical solution of the present application, the transmission between the driving mechanism and the main shaft of the eccentric shaft is belt transmission.

[0025] As an improved technical solution of the present application, it also includes a base, which is located below the point shock table and has a through hole for the point shock rod to pass through.

[0026] As an improved technical solution of the present application, it also includes a point shock rod protective cover, which is coaxially installed with the point shock rod and fixed to the lower surface of the base.

[0027] As an improved technical solution of the present application, the point vibration table is provided with limit pins at the corners.

[0028] Beneficial Effects

[0029] The present application uses a method of tapping the cell culture container perpendicularly to its bottom surface to achieve efficient digestion of adherent cells into single suspended cells; compared with the prior art that uses centrifugal force vibration (point vibration) generated by horizontal eccentric rotation to culture adherent cells, from the analysis of actual application effects, the viability of adherent cultured cells after digestion can be increased by more than 20% and the agglomeration rate can be reduced by 30%, especially for the digestion of adherent cells cultured on a large scale.

[0030] The technical solution of the present application is suitable for the culture of adherent cells, including but not limited to the use of large-scale cell culture containers such as single-layer or multi-layer cell factories for large-scale culture of adherent cells.

[0031] The technical solution of the present application can be automatically controlled and can cooperate with large-scale unmanned automated production lines to ensure consistency between batches for commercial application.

[0032] The elastomer of the present application is embedded in the point vibration through hole, meeting the GMP environmental requirements for culturing cells; at the same time, the elastomer is easy to install and convenient to replace, clean and maintain. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 A schematic diagram of the structure of the point vibration table of the present application;

[0034] Figure 2 A schematic diagram of the structure of the through hole of the point vibration platform of the present application;

[0035] Figure 3 A side view of the earthquake station at the point of application;

[0036] Figure 4 The front view of the three-dimensional structure of the point seismic station of this application;

[0037] Figure 5 A schematic diagram of the back side of the three-dimensional structure of the point seismic station of the present application;

[0038] Figure 6 Schematic diagram of the connection relationship of the point shock rod;

[0039] Figure 7 A schematic diagram of the structure of the driving mechanism of this application;

[0040] Figure 8 Schematic diagram of the eccentric shaft structure of this application;

[0041] In the figure: 1. point shock table; 2. point shock through hole; 3. limit pin; 4. base; 5. point shock rod protective cover; 6. first connecting piece; 7. belt; 8. driving mechanism; 9. support plate; 10. eccentric shaft; 101. main shaft; 102. output shaft; 11. point shock rod; 12. second connecting piece; 13. point shock point; 14. back support plate; 15. point shock rod support body; 16. connecting rod; 17. elastic body. DETAILED DESCRIPTION

[0042] definition:

[0043] Point vibration table top: a table top with point vibration function. Point vibration means the ability to achieve regular impact on a fixed point or fixed plane.

[0044] A connecting rod mechanism is a mechanism in which multiple connecting rods are assembled in a specific way, which can change the direction of force from one direction to another.

[0045] Point shock point 13: the contact point where the shock rod 11 performs point shock on the elastic body 17.

[0046] Adherent cells refer to cells that must have a support surface to attach to when growing. The cells rely on the attachment factors secreted by themselves or provided in the culture medium to grow and reproduce on the surface. They include cells that can attach and grow on culture containers (for example, culture dishes, culture bottles, cell factories, etc.) or other solid surfaces.

[0047] Lower surface: the side of the large-scale cell culture container facing away from the point-vibration table 1.

[0048] The present application provides a point vibration table suitable for culturing adherent cells, which is suitable for digesting and dispersing adherent cells cultured in large-scale cell culture containers such as cell factories. The present application converts the eccentric motion of the eccentric shaft 10 into vertical motion, performs point vibration impact on the bottom surface of the cell culture container, and causes the adherent cells to detach and disperse from the bottom surface of the cell culture container. Compared with the prior art such as oscillators, which mainly use centrifugal force and vortex force to make the sample rotate rapidly in the container, thereby generating strong convection and shear force to promote cell dispersion, the technical solution of the present application causes minimal damage to the cells and can achieve efficient dispersion of adherent cells cultured in large-scale culture containers.

[0049] In particular, in order to reduce pollution, the technical solution of the present application is designed with an elastic body 17. The point shock impact on the large-scale cell culture container is: the eccentric movement of the eccentric shaft 10 is converted into vertical movement to perform point shock impact on the elastic body 17. This method can not only avoid direct impact on the bottom surface of the cell culture container to cause cell damage or damage to the cell culture container; but also isolate the static space of the cell culture container from the space with dynamic movement such as the eccentric shaft 10, the connecting rod mechanism and the driving mechanism 8, so as to avoid the damage of the environment of the cell culture container by the particles generated during the movement of the eccentric shaft 10, the connecting rod mechanism and the driving mechanism 8.

[0050] The technical solution of this application will be clearly and completely described below in conjunction with the accompanying drawings.

[0051] A point vibration table suitable for culturing adherent cells comprises a point vibration table surface 1, an elastic body 17, an eccentric shaft 10, a connecting rod mechanism and a driving mechanism 8.

[0052] (I) Point vibration table 1

[0053] In actual application, the point vibration table 1 mainly supports the cell culture container and the elastic body 17 to rise and fall, and the point vibration table 1 itself is not vibrated by the eccentric shaft 10 and the connecting rod mechanism. The mechanical properties of the point vibration table 1 can support the elastic body 17 to be fully point-vibrated and not fall off from the point vibration through hole 2. In some embodiments, the point vibration table 1 is a plane.

[0054] The point vibration table 1 has a point vibration through hole 2 on its upper surface. The arrangement position of the point vibration through hole 2 matches the shape and size of the cell culture container; if a rectangular cell factory is used as a cell container to complete the large-scale culture of adherent cells, the layout position of the point vibration through hole 2 is designed in combination with the rectangular shape and size of the cell factory.

[0055] In some embodiments, one spot-vibration table 1 performs spot-vibration on the bottom surface of only one culture container.

[0056] Embodiment 1: A point-vibration table 1 is provided with only one point-vibration through hole 2 to perform point-vibration on a culture container.

[0057] This method is based on the cooperation of a point-vibration through hole 2, an elastic body 17, a connecting rod mechanism, an eccentric shaft 10 and a driving mechanism 8 to achieve point-vibration. Specifically, a point-vibration through hole 2 is used to point-vibrate the bottom surface of a culture container; an elastic body 17 is embedded in the point-vibration through hole 2, and the elastic body 17 is point-vibrated by a point-vibration rod 11, and the point-vibration rod 11 has only one point-vibration point 13; the point-vibration rod 11 is driven by the connecting rod mechanism to perform linear motion.

[0058] Embodiment 2: A point-vibration table 1 is provided with a group of point-vibration through holes 2 (a plurality of point-vibration through holes 2) so as to perform point-vibration on a culture container at the same time.

[0059] This method is based on the cooperation of multiple point-vibration through holes 2, multiple elastic bodies 17 (a group of elastic bodies 17), a connecting rod mechanism, an eccentric shaft 10 and a driving mechanism 8 to achieve point-vibration. Specifically, multiple point-vibration through holes 2 are used to perform point-vibration on the bottom surface of a culture container; multiple elastic bodies 17 are correspondingly embedded and installed in multiple point-vibration through holes 2, and a point-vibration rod with multiple point-vibration points 13 is selected, and the multiple point-vibration points 13 correspond to the multiple elastic bodies 17 one by one. The point-vibration rod 11 with multiple point-vibration points 13 is driven by the connecting rod mechanism to perform linear motion to achieve simultaneous point-vibration of multiple elastic bodies 17.

[0060] In some embodiments, one spot-vibration table 1 performs spot-vibration on the bottom surfaces of only two culture containers.

[0061] In the third embodiment, a point-vibration table 1 performs point-vibration on the bottom surfaces of two culture containers at the same time through two point-vibration through holes (each point-vibration through hole 2 performs point-vibration on the bottom surface of one culture container). The difference from the first embodiment is that: there are two elastic bodies 17, and each elastic body 17 is correspondingly embedded and installed in one point-vibration through hole 2; the eccentric shaft 10 includes a main shaft and two output shafts located at both ends of the main shaft; the axes of the two output shafts are respectively parallel to the axis of the main shaft, and are axially symmetrical with respect to the main shaft; each output shaft is connected to one of the connecting rod mechanisms; each of the connecting rod mechanisms performs point-vibration on one elastic body 17.

[0062] Implementation method 4: Figure 2 As shown, a point vibration table 1 simultaneously performs point vibration on the bottom surfaces of two culture containers through two groups of point vibration through holes 2 (each group of point vibration through holes 2 performs point vibration on the bottom surface of one culture container). Figure 1 and Figure 7As shown, what is different from the second embodiment is that: the point shock table 1 is provided with two groups of point shock through holes 2; each group of point shock through holes 2 is composed of a plurality of point shock through holes 2; there are two groups of elastomers 17, and the number of elastomers 17 in each group corresponds to the number of point shock through holes 2 in each group; the eccentric shaft 10 includes a main shaft and two output shafts located at both ends of the main shaft; the axes of the two output shafts are respectively parallel to the axis of the main shaft, and are axially symmetrical with respect to the main shaft; each output shaft is connected to one of the connecting rod mechanisms; each of the connecting rod mechanisms drives the point shock rod 11 to perform point shock on a group of elastomers 17.

[0063] In the above-mentioned second and fourth embodiments, Figure 6 As shown, the point shock rod includes a plurality of point shock points 13, which correspond one to one with a group of elastic bodies 17;

[0064] A connecting rod 16 connects all the point-vibration points 13 so that all the point-vibration points 13 move linearly at the same time;

[0065] The shock rod support body 15 has one end driven by the output shaft through the connecting rod mechanism and the other end supporting the connecting rod 16 .

[0066] In order to ensure the stability of the cell culture container, the point vibration table 1 is provided with limit pins 3 at the corners, specifically, the limit pins 3 are provided at the four corners of the point vibration table 1 near the point vibration through hole 2. The function of the limit pins 3 is to ensure that the container for large-scale cell culture can be stably present on the point vibration table 1.

[0067] (ii) Elastic body 17

[0068] The elastic body 17 has a groove on its side that matches the point shock through hole 2; the elastic body 17 is embedded in the point shock through hole 2, and the groove accommodates the periphery of the point shock through hole 2. In this technical solution, the material selection requirement for the elastic body 17 is: to meet the safety of biological products. Therefore, in some embodiments, the elastic body 17 is a silicone elastic body 17; in other embodiments, the elastic body 17 can also be a polyurethane elastomer 17, etc.

[0069] (three) eccentric shaft 10

[0070] like Figure 8 As shown, the eccentric shaft 10 includes a main shaft 101 and an output shaft 102, and the output shaft 102 is not coaxial with the main shaft 101; that is, when the driving mechanism 8 is working, the axis of the output shaft 102 is parallel to the axis of the main shaft 101 but does not overlap. The main purpose is to utilize the eccentric setting of the output shaft 102 relative to the main shaft 101, and the rotation of the main shaft 101 drives the output shaft 102 to rotate, thereby generating an eccentric force, which is the original force providing the point vibration effect.

[0071] Figure 8 The eccentric shaft 10 shown has two output shafts 102 to perform point vibration on two point vibration through holes or two groups of point vibration through holes. In actual application, only one output shaft 102 can be designed to perform point vibration on one point vibration through hole.

[0072] (IV) Connecting rod mechanism

[0073] like Figure 3-Figure 5 The function of the connecting rod mechanism in the present application is to convert the circumferential rotation of the output shaft of the eccentric shaft 10 into a linear motion that causes the shock rod 11 to move perpendicularly to the lower surface of the elastic body 17 .

[0074] The installation method is as follows: one end is installed on the output shaft through a bearing, and the other end is installed with a point shock rod 11, and the point shock rod 11 is located on the lower surface of the elastic body 17; at the same time, when the main shaft rotates, the point shock rod 11 only moves linearly perpendicular to the lower surface of the elastic body 17, and performs point shock on the elastic body 17. The connecting rod mechanism can be any connecting structure in the prior art that can change the direction of force, such as an eccentric wheel linear reciprocating mechanism, etc.

[0075] Due to the installation position, the direction of linear motion and other reasons, the connecting rod mechanism of this application is designed to include:

[0076] The first connecting member 6 is an L-shaped structure, and a first mounting hole and a second mounting hole are respectively provided at both ends of the L-shaped structure; the purpose of the L-shaped structure is to change the direction of the eccentric force of the eccentric shaft 10; the second connecting member 12 is provided with a first bearing mounting hole at one end and a second bearing mounting hole at the other end; a bearing is installed in the second bearing mounting hole, and the output shaft of the eccentric shaft 10 is installed in the first bearing mounting hole; another bearing is installed in the second bearing mounting hole, and the first mounting hole and the second bearing mounting hole are connected by a connecting shaft;

[0077] One end of the point shock rod 11 is fixed on the second mounting hole and supported by the second mounting hole; the other end is the point shock point 13 for vibrating the elastic body 17.

[0078] (V) Point shock rod 11

[0079] In some embodiments, the shock rod 11 includes a shock point 13 and a shock rod support 15, wherein the shock point 13 is located at the end of the shock rod support 15, and the shock rod support 15 is driven by the output shaft through the connecting rod mechanism.

[0080] In other embodiments, the shock rod 11 includes a plurality of shock points 13 corresponding to each group of multiple elastic bodies 17; a connecting rod 16 connecting all shock points 13 so that all shock points 13 move linearly at the same time; and a shock rod support 15, one end of which is driven by the output shaft through the connecting rod mechanism and the other end supports the connecting rod 16. Specifically, the shock rod 11 in Embodiment 2 and Embodiment 4.

[0081] (VI) Driving mechanism 8

[0082] The driving mechanism 8 drives the main shaft of the eccentric shaft 10 to rotate. The driving mechanism 8 mainly adopts a motor. The transmission between the driving mechanism 8 and the main shaft of the eccentric shaft 10 is a belt 7 transmission.

[0083] (7) Auxiliary components

[0084] In practical applications, a base 4 is provided below the point shock table 1, and a through hole is provided on the base 4 for the point shock rod 11 to pass through; at the same time, a point shock rod protective cover 5 is provided at the position of the through hole for the point shock rod 11 to pass through. That is, the point shock rod protective cover 5 is coaxially installed with the point shock rod 11 and fixed to the lower surface of the base 4.

[0085] The bottom surface of the base 4 is also provided with a support plate 9, on which a drive mechanism 8 mounting hole and an eccentric shaft 10 through hole are opened; the drive mechanism 8 is installed on the drive mechanism 8 mounting hole, and the main shaft of the eccentric shaft 10 is installed at the eccentric shaft 10 mounting through hole.

[0086] The back support plate 14 supports the base 4 and the support plate 9 at the same time, thereby promoting the overall stability of the point vibration platform.

Claims

1. A point vibration platform suitable for culturing adherent cells, characterized in that: include The point-vibration table is provided with a point-vibration through hole; An elastic body, a groove matching the point-vibration through hole is provided on its side; the elastic body is embedded in the point-vibration through hole, and the groove accommodates the periphery of the point-vibration through hole; An eccentric shaft comprises a main shaft and an output shaft, wherein the output shaft is not coaxial with the main shaft; A connecting rod mechanism, one end of which is mounted on the output shaft through a bearing, and the other end of which is mounted with a point-shock rod; the point-shock rod is located on the lower surface of the elastic body; when the main shaft rotates, the point-shock rod only moves linearly perpendicular to the lower surface of the elastic body and performs point-shock on the elastic body; A driving mechanism drives the main shaft of the eccentric shaft to rotate.

2. A point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: The point vibration table has two point vibration through holes; there are two elastomers, each of which is embedded and installed in a corresponding point vibration through hole; the eccentric shaft includes a main shaft and output shafts located at both ends of the main shaft; the axes of the two output shafts are respectively parallel to the axis of the main shaft and are axially symmetrical with respect to the main shaft; each output shaft is connected to a connecting rod mechanism; each connecting rod mechanism performs point vibration on one of the elastomers.

3. A point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: The point shock table is provided with two groups of point shock through holes; each group of point shock through holes is composed of a plurality of point shock through holes; there are two groups of elastomers, and the number of elastomers in each group corresponds to the number of point shock through holes in each group; the eccentric shaft includes a main shaft and output shafts located at both ends of the main shaft; the axes of the two output shafts are parallel to the axes of the main shaft, and are axially symmetrical relative to the main shaft; each output shaft is connected to a connecting rod mechanism; the point shock rod in each connecting rod mechanism contains a plurality of point shock points, and the number of point shock points in each connecting rod mechanism corresponds to a group of elastomers; the point shock points complete the point shock of the elastomer.

4. A point vibration platform suitable for culturing adherent cells according to claim 3, characterized in that: The point shock rod comprises There are multiple point shock points, which correspond one by one to a group of elastic bodies; A connecting rod connects all the point-vibration points so that all the point-vibration points move linearly at the same time; A point shock rod support body, one end of which is driven by the output shaft through the connecting rod mechanism, and the other end of which supports the connecting rod.

5. The point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: The connecting rod mechanism comprises The first connecting member is in an L-shaped structure, and a first mounting hole and a second mounting hole are respectively provided at two ends of the L-shaped structure; A second connecting member, one end of which is provided with a first bearing mounting hole, and the other end of which is provided with a second bearing mounting hole; a bearing is installed in the first bearing mounting hole, and the output shaft of the eccentric shaft is installed in the first bearing mounting hole; another bearing is installed in the second bearing mounting hole, and the first mounting hole and the second bearing mounting hole are connected by a connecting shaft; One end of the point shock rod is fixed on the second mounting hole and supported by the second mounting hole; the other end is a point shock point for vibrating the elastic body.

6. A point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: The elastic body is made of silicone.

7. A point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: The transmission between the driving mechanism and the main shaft of the eccentric shaft is belt transmission.

8. The point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: It also includes a base, which is located below the point shock table and is provided with a through hole for the point shock rod to pass through.

9. A point vibration platform suitable for culturing adherent cells according to claim 8, characterized in that: It also includes a shock rod protection cover, which is coaxially installed with the shock rod and fixed to the lower surface of the base.

10. The point vibration platform suitable for culturing adherent cells according to claim 1, characterized in that: The point vibration table is provided with limiting pins at the corners.

Images