CN114369533A - Animal cell tissue culture device and culture method - Google Patents
Animal cell tissue culture device and culture method Download PDFInfo
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- CN114369533A CN114369533A CN202210033187.XA CN202210033187A CN114369533A CN 114369533 A CN114369533 A CN 114369533A CN 202210033187 A CN202210033187 A CN 202210033187A CN 114369533 A CN114369533 A CN 114369533A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
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- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/16—Vibrating; Shaking; Tilting
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/10—Perfusion
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
Abstract
The invention relates to the technical field of animal cell tissue culture, in particular to an animal cell tissue culture device and a culture method, wherein the device comprises a culture platform, a connecting column, a culture frame, a cover plate, a shaking mechanism, a bearing plate, a culture dish and a feeding mechanism, wherein in the shaking mechanism, a rotating disc drives a vertical moving block to move up and down in a reciprocating manner on an annular adjusting plate, and then the vertical moving block drives the bearing plate to synchronously drive the bearing plate to shake in the rotating process through a shaking rod, so that nutrient solution in the culture dish is shaken, shaken and evenly shaken; according to the amount of required nutrient solution in each culture dish in feed mechanism, open the valve and pass through injection cylinder extrusion air with the nutrient solution through liquid distribution pipe and connecting pipe mutually support with the nutrient solution pump income culture dish, do not open the apron at whole material loading in-process to reduce the possibility that miscellaneous fungus got into the cavity inside, reduce the possibility that miscellaneous fungus pollutes animal tissue cell culture.
Description
Technical Field
The invention relates to the technical field of animal cell tissue culture, in particular to an animal cell tissue culture device and a culture method.
Background
Animal cell culture is a method for obtaining new cells by taking relevant tissues from animals and placing the tissues in a culture solution to allow the cells of the tissues to grow and proliferate, and with the development of gene engineering and other cell engineering technologies, the cell culture technology has become the basis of transgenic technology, biopharmaceutical technology and other various technologies, and plays an important role in modern biotechnology.
Animal tissue cells scraped by a scraper are placed in the middle of a culture dish manually, and because the animal tissue cells need to consume nutrient components in a culture solution in the proliferation and growth process, the concentration of the nutrient components in the culture solution is gradually reduced from the middle of the culture dish to the periphery of the culture dish in the culture process of the animal tissue cells, and the nutrient components in the culture solution have the problem of non-uniformity, so that the proliferation effect of the animal tissue cells is influenced.
Because animal tissue cell probably needs in time to supply the culture solution at the culture in-process, need artifically open culture apparatus and directly add the culture solution to the culture dish in traditional culture solution supply in-process, this in-process can cause the mixed fungus in the air to enter into the culture dish, leads to mixed fungus to enter into the culture solution and influences animal tissue cell growth, and harmful mixed fungus even can cause animal tissue cell proliferation to stop, influences animal tissue cell culture result.
Disclosure of Invention
In order to solve the problems, the invention adopts the following technical scheme: the utility model provides an animal cell tissue culture device, includes culture platform, spliced pole, culture frame, apron, rocks mechanism, bearing dish, culture dish and feed mechanism, four corners of culture platform up end install the spliced pole, four spliced pole up ends install culture frame jointly, four cavities are separated into through the cross baffle to culture frame inside, rock the mechanism inside and culture platform up end install jointly to rock four cavities, rock the mechanism upper end and install bearing dish, the joint has the culture dish on the bearing dish, culture frame up end is installed the apron, the feed mechanism is installed to the apron up end, the apron up end just is located the cavity and installs the micro-magnifier of observing the cavity directly over, the observation window has been seted up to four corners of culture frame lateral wall.
The shaking mechanism comprises a shaking motor, a rotating shaft, a rotating disc, an annular adjusting plate, vertical moving blocks, vertical connecting shafts, a shaking rod and a rotating sleeve, wherein the shaking motor is arranged in the middle of the upper end face of the culture platform through a motor base, the rotating shaft is arranged at the lower end of the culture frame and positioned in the middle of four cavities in a penetrating way through bearings, the output shaft of the shaking motor is mutually connected with the four rotating shafts in a gear meshing way, the rotating disc is arranged on the upper end face of the rotating shaft, the annular adjusting plate is arranged in the middle of the bottom of the cavity, a plurality of adjusting grooves are evenly arranged on the upper end face of the annular adjusting plate in the circumferential direction, the vertical moving blocks are arranged on the upper end of the rotating disc in a penetrating and sliding way up and down, the lower ends of the vertical moving blocks abut against the annular adjusting plate, the vertical connecting shafts are arranged on the middle of the upper end face of the rotating disc through bearings, and a bearing tray is arranged on the upper end face of the vertical connecting shafts through ball hinges, the middle part of the vertical connecting shaft is provided with a through groove from left to right, a rotating sleeve is installed inside the through groove through a hinge shaft, a shaking rod is arranged inside the rotating sleeve in a sliding mode, a moving connecting block is arranged on the lower end face of the bearing plate and the upper end face of the vertical moving block in a sliding mode, the upper end of the shaking rod is installed on the moving connecting block above the supporting plate through a hinge, and the lower end of the shaking rod is installed on the moving connecting block below the shaking rod through a hinge.
Feed mechanism include the material loading post, go up the feed cylinder, injection tube, liquid distribution pipe and connecting pipe, wherein the material loading post install terminal surface middle part on the apron, it installs in the material loading post upper end through the threaded connection mode to go up the feed cylinder, go up the feed inlet of installing the sealing plug on the left of the feed cylinder, go up the feed cylinder up end and install and link up mutually with the material loading cylinder and have the injection tube of scale, a plurality of liquid distribution pipes that link up mutually with last feed cylinder are evenly installed to terminal surface circumference under the material loading cylinder, also install the liquid distribution pipe that link up mutually with the cavity directly over the apron up end and being located four culture dishes, the joint has the connecting pipe that has the valve between two upper and lower adjacent liquid distribution pipes.
Preferably, the middle part of the lower end face in the upper charging barrel is provided with a rotating shaft through a bearing, a plurality of stirring claws are uniformly arranged below the circumferential surface of the rotating shaft, a first bevel gear is arranged above the rotating shaft, a linkage shaft is arranged above the right side wall of the upper charging barrel through a bearing, a second bevel gear meshed with the first bevel gear is arranged on the left end face of the linkage shaft, a transmission gear is arranged on the linkage shaft, the upper end of the upper charging barrel is provided with a sliding plate in a penetrating mode from top to bottom, the upper end of the sliding plate is abutted against the lower end face of the telescopic end of an injection tube, the lower end of the front end face of the sliding plate is provided with a rack matched with the transmission gear, the bottom of the sliding plate is provided with a horizontal fixing bulge, and a reset spring rod is arranged between the sliding plate and the inner wall of the upper charging barrel through the fixing bulge.
Preferably, four the cavity inner wall and be located the bearing dish outside and all install the loop forming element, and the loop forming element sets up the structure that the groove was led to the circle for square board up end, evenly install a plurality of bearing spring levers on the loop forming element inner wall, the one end that the loop forming element was kept away from to the bearing spring lever is installed through the ball pivot and is born tray matched with bearing piece, the loop forming element can play spacing bearing effect to the bearing dish through bearing spring lever and bearing piece mutually supporting, avoid bearing the dish at vertical epaxial single-point atress instability of company, increase and bear the tray and rock the in-process stable effect.
Preferably, the inside evenly set up a plurality of guide plates of seting up the guide hole of connecting pipe, the drainage plate is installed through the support frame in the branch liquid pipe exit that terminal surface just is located the below under the apron, the drainage plate up end sets up to the toper structure, the drainage plate lower extreme sets up to the umbrella type structure, umbrella type structure up end circumference evenly is provided with a plurality of drainage grooves, the lower extreme is umbrella type structure's drainage plate can play the water conservancy diversion effect to the culture solution through mutually supporting with the drainage groove, it is inhomogeneous to avoid the culture solution to distribute when getting into the culture dish, cause the inhomogeneous problem of animal tissue cell acquisition nutrient composition, influence the proliferation effect of animal tissue cell, the guide plate can reduce the impact force that the culture solution got into the culture dish, avoid too big impact force of culture solution to break away animal tissue cell, influence the culture condition of animal tissue cell.
Preferably, the connecting springs are symmetrically arranged between the guide plates and the inner wall of the connecting pipe from top to bottom, the wiping sponge is arranged at the contact part of the guide plates and the inner wall of the connecting pipe, the guide rope is arranged between two adjacent guide plates on the same side of the valve, and the diameter of the connecting pipe is small and long, so that the guide rope is manually driven to drive the guide plates to slide in the connecting pipe in a reciprocating manner in the water inlet cleaning process of the connecting pipe, the inner wall of the connecting pipe can be cleaned in a reciprocating manner through the wiping sponge by the guide plates, the cleaning of the inside of the connecting pipe is facilitated, and the connecting springs play a role in resetting the cleaning process of the guide plates.
Preferably, apron lower extreme face edge be provided with the rectangle arch, cultivate frame upper end face edge and seted up with rectangle arch matched with seal groove, the seal groove mutually supports with the rectangle arch can the cavity inside play sealed effect, avoids the impurity in the air to enter into the cavity inside, influences the animal tissue cell growth condition.
Preferably, the inner wall of the culture dish on the annular plate is clamped, the annular plate is clamped on the inner wall of the culture dish after the addition of the animal tissue cells and the culture solution in the culture dish is finished, the phenomenon that the culture solution splashes out of the culture dish in the sliding process of the culture dish to cause the waste of the culture solution is avoided, and the sanitation inside the cavity is ensured.
Preferably, the adjusting grooves are different in depth, so that the vertical moving block obtains different amplitudes in the up-and-down moving process, the shaking rod drives the bearing plate to obtain different shaking amplitudes, and therefore the culture dish is guaranteed to obtain different shaking amplitudes, and nutrient components in the culture solution are distributed more uniformly.
Preferably, the method for culturing animal cells by using the animal cell tissue culture device comprises the following steps: the first step is as follows: placing treatment: adding culture solution into the culture dish filled with the animal tissue cells, taking down the cover plate, then clamping the culture dish inside the bearing tray, and finally closing the cover plate.
The second step is that: shaking treatment: the shaking motor is started to drive the rotating shaft to rotate in a gear transmission mode, the rotating shaft drives the vertical moving block to reciprocate up and down on the annular adjusting plate through the rotating disc, and then the vertical moving block drives the bearing plate to synchronously drive the bearing plate to shake in the rotating process of the bearing plate through the shaking rod, so that the culture solution in the culture dish is shaken and evenly shaken.
The third step: and (3) loading treatment: when the culture solution in the culture dish is not enough, open the sealing plug and go up the inside culture solution of pouring into of feed inlet to the feed cylinder, close the sealing plug again, according to how much of required culture solution in each culture dish, open the valve and pass through injection tube extrusion air with the culture solution through the branch liquid pipe with the connecting pipe mutually support add the culture solution in the culture dish.
The fourth step: and (4) observation and taking out: observing the growth condition of the animal tissue cells through a microscopic magnifier, opening the cover plate to take out the culture dish after the culture of the animal tissue cells is finished, and finishing the work.
The invention has the beneficial effects that: 1. the culture frame designed by the invention is divided into four cavities, so that the animal cell tissues in four culture dishes can be subjected to proliferation culture simultaneously in the process of subculture of the animal cell tissues, and the proliferation culture efficiency of the animal cell tissues is improved; in the shaking mechanism, the rotating disc drives the vertical moving block to reciprocate up and down on the annular adjusting plate, and then the vertical moving block drives the bearing plate to shake in the rotating process of the bearing plate through the shaking rod, so that the culture solution in the culture dish is subjected to shaking and shaking uniformly; in feed mechanism, open the sealing plug and go up the inside culture solution of pouring into of feed inlet to the feed cylinder, close the sealing plug again, according to how much of required culture solution in each culture dish, open the valve and pass through injection tube extrusion air with the culture solution through the branch liquid pipe with the connecting pipe mutually support with the culture solution pump in the culture dish, do not open the apron at whole material loading in-process to reduce the possibility that miscellaneous fungus got into the cavity is inside, reduce the possibility that miscellaneous fungus pollutes animal tissue cell culture.
2. In the feeding mechanism designed by the invention, when the culture solution required in each culture dish is the same, the valve is synchronously opened, the culture solution is mutually matched with the connecting pipe through the liquid distribution pipe by extruding air through the injection pipe, the culture solution is pumped into the culture dish, when the culture solution required in each culture dish is different, the valve is respectively opened, the culture solution is mutually matched with the connecting pipe through the liquid distribution pipe by extruding air through the injection pipe, the culture solution is pumped into the culture dish, the culture solution can be synchronously pumped into the culture dish and can also be respectively pumped into the culture dish by adjusting the valve, and the applicability is wider.
3. In the feeding cylinder designed by the invention, when the telescopic end of the injection cylinder is pushed to move downwards, the sliding plate synchronously moves downwards, the sliding plate drives the transmission gear to rotate through the rack, the transmission gear drives the linkage shaft to rotate, the linkage shaft drives the rotating shaft to rotate through the mutual matching of the first bevel gear and the second bevel gear, so that the rotating shaft drives the stirring claw to uniformly stir the culture solution, the problem that the nutrient components are likely to precipitate due to the fact that the culture solution is in a standing state for a long time after being filled into the feeding cylinder is avoided, and the uniformity of nutrient substances in the culture solution is improved.
Drawings
The invention is further illustrated with reference to the following figures and examples.
Fig. 1 is a perspective view of the present invention.
Fig. 2 is a front view of the present invention.
FIG. 3 is a schematic perspective view of the inside of the culture frame of the present invention.
FIG. 4 is a schematic view of the internal installation structure of the cavity and the rocking mechanism of the present invention.
Fig. 5 is a partial cross-sectional view at a of fig. 4 of the present invention.
FIG. 6 is a schematic view of a three-dimensional mounting structure between the cavity and the rocking mechanism of the present invention.
Fig. 7 is a schematic view of the internal structure of the charging barrel of the invention.
FIG. 8 is a schematic view of the internal structure of the connecting tube of the present invention.
Fig. 9 is a partial enlarged view of the invention at B of fig. 8.
FIG. 10 is a perspective view of a flow guide plate according to the present invention.
FIG. 11 is a flow chart of tissue culture of animal cells according to the present invention.
In the figure, 1, culture platform; 2. connecting columns; 3. a culture frame; 31. a sealing groove; 32. a rectangular protrusion; 4. a cover plate; 41. a microscopic magnifier; 42. an observation window; 5. a shaking mechanism; 51. shaking the motor; 52. a rotating shaft; 53. rotating the disc; 54. an annular adjusting plate; 541. adjusting the groove; 55. a vertical moving block; 56. a vertical connecting shaft; 561. a through groove; 57. a shaking rod; 58. rotating the sleeve; 59. moving the connecting block; 6. a support tray; 61. an annular member; 62. supporting the spring rod; 63. a bearing block; 7. a culture dish; 71. an annular plate; 8. a feeding mechanism; 81. feeding a material column; 82. feeding a material barrel; 821. a rotating shaft; 822. a stirring claw; 823. a first bevel gear; 824. a linkage shaft; 825. a second bevel gear; 826. a transmission gear; 827. a sliding plate; 828. a return spring lever; 83. an injection tube; 84. a liquid separating pipe; 85. a connecting pipe; 851. a guide plate; 852. a drainage plate; 853. a drainage groove; 854. a connecting spring; 855. wiping sponge; 856. a guide rope; 86. a valve; 87. and (4) sealing the plug.
Detailed Description
The embodiments of the invention will be described in detail below with reference to the drawings, but the invention can be implemented in many different ways as defined and covered by the claims.
As shown in FIG. 1, FIG. 3 and FIG. 4, an animal cell tissue culture device comprises a culture platform 1, a connecting column 2, a culture frame 3, a cover plate 4, a shaking mechanism 5, a support tray 6, a culture dish 7 and a feeding mechanism 8, four corners of 1 up end of culture platform install spliced pole 2, culture frame 3 is installed jointly to four 2 up ends of spliced pole, culture frame 3 is inside separates into four cavitys through the cross baffle, four cavitys are inside and culture platform 1 up end is installed jointly and is rocked mechanism 5, rock mechanism 5 upper end and install bearing tray 6, the joint has culture dish 7 on the bearing tray 6, apron 4 is installed to culture frame 3 up end, feed mechanism 8 is installed to apron 4 up end, apron 4 up end just is located the cavity and installs the micro-magnifier 41 who observes the cavity directly over, observation window 42 has been seted up to four corners of culture frame 3 lateral walls.
As shown in fig. 3 and 4, the edge of the lower end face of the cover plate 4 is provided with a rectangular protrusion 32, the edge of the upper end face of the culture frame 3 is provided with a sealing groove 31 matched with the rectangular protrusion 32, the sealing groove 31 and the rectangular protrusion 32 are matched with each other to seal the inside of the cavity, and the situation that impurities in the air enter the cavity to affect the growth of animal tissue cells is avoided.
As shown in fig. 6, the inner wall of the culture dish 7 is connected with the annular plate 71 in a clamped manner, and the annular plate 71 is connected to the inner wall of the culture dish 7 after the addition of the animal tissue cells and the culture solution in the culture dish 7 is finished, so that the culture dish 7 is prevented from splashing out of the culture solution from the culture dish 7 in the shaking process, the waste of the culture solution is avoided, and the sanitation inside the cavity is ensured.
As shown in fig. 2, 4, 5 and 6, the shaking mechanism 5 comprises a shaking motor 51, a rotating shaft 52, a rotating disc 53, an annular adjusting plate 54, a vertical moving block 55, a vertical connecting shaft 56, a shaking rod 57 and a rotating sleeve 58, wherein the shaking motor 51 is installed in the middle of the upper end surface of the culture platform 1 through a motor base, the rotating shaft 52 is installed at the lower end of the culture frame 3 and positioned in the middle of four cavities through bearings in a penetrating manner, the output shaft of the shaking motor 51 and the four rotating shafts 52 are all connected with each other through a gear meshing manner, the rotating disc 53 is installed on the upper end surface of the rotating shaft 52, the annular adjusting plate 54 is installed at the middle position of the bottom of the cavity, a plurality of adjusting grooves 541 are evenly formed in the circumferential direction of the upper end surface of the annular adjusting plate 54, the vertical moving block 55 is installed on the upper end of the rotating disc 53 in a penetrating and sliding manner, and the lower end of the vertical moving block 55 is abutted against the annular adjusting plate 54, the middle of the upper end face of the rotating disc 53 is provided with a vertical connecting shaft 56 through a bearing, the upper end face of the vertical connecting shaft 56 is provided with a bearing tray 6 through a spherical hinge, the middle of the vertical connecting shaft 56 is provided with a through groove 561 from left to right, a rotating sleeve 58 is arranged in the through groove 561 through a hinge shaft, a shaking rod 57 is arranged in the rotating sleeve 58 in a sliding mode, the lower end face of the bearing tray 6 and the upper end face of the vertical moving block 55 are respectively provided with a moving connecting block 59 in a horizontal sliding mode, the upper end of the shaking rod 57 is arranged on the moving connecting block 59 above through a hinge, and the lower end of the shaking rod 57 is arranged on the moving connecting block 59 below through a hinge.
As shown in fig. 4 and 6, the depths of the adjacent adjusting grooves 541 are different, so that the vertical moving block 55 obtains different amplitudes during the up-and-down movement, and then the shaking rod 57 drives the supporting plate 6 to obtain different shaking amplitudes, thereby ensuring that the culture dish 7 obtains different shaking amplitudes and the nutrient content inside the culture solution is more uniformly distributed.
As shown in fig. 1, the feeding mechanism 8 includes a feeding column 81, a feeding cylinder 82, an injection tube 83, a liquid distribution tube 84 and a connecting tube 85, wherein the feeding column 81 is installed in the middle of the upper end surface of the cover plate 4, the feeding cylinder 82 is installed at the upper end of the feeding column 81 through a threaded connection mode, a feeding hole for installing a sealing plug 87 is opened at the left side of the feeding cylinder 82, the injection tube 83 which is communicated with the feeding cylinder 82 and has scale is installed on the upper end surface of the feeding cylinder 82, a plurality of liquid distribution tubes 84 which are communicated with the feeding cylinder 82 are evenly installed on the lower end surface of the feeding cylinder 82 in the circumferential direction, the liquid distribution tube 84 which is communicated with the cavity is also installed right above the four culture dishes 7 on the upper end surface of the cover plate 4, and the connecting tube 85 with a valve 86 is clamped between the upper and lower two adjacent liquid distribution tubes 84.
During the specific operation, the culture solution is manually added into the culture dish 7 containing the animal tissue cells, the cover plate 4 is taken down and then the culture dish 7 is clamped inside the bearing tray 6, finally the cover plate 4 is closed, the animal tissue cells proliferate and grow inside the culture dish 7, the growth condition of the animal tissue cells is observed through the micro magnifier 41 and the observation window 42, as the animal tissue cells need to consume nutrient components in the culture solution during the proliferation and growth processes, the concentration of the nutrient components in the culture solution is gradually reduced from the middle of the culture dish 7 to the periphery of the culture dish 7, so that the nutrient components in the culture solution are uneven, the growth of the animal tissue cells is influenced, therefore, the shaking motor 51 is started to drive the rotating shaft 52 to rotate through a gear meshing mode in the culture process of the animal tissue cells, the rotating shaft 52 drives the vertical moving block 55 to synchronously rotate through the rotating disc 53, the vertical moving block 55 drives the bearing tray 6 to rotate around the vertical connecting shaft 56 through the shaking rod 57, the vertical moving block 55 in the rotating process of the supporting plate 6 drives the shaking rod 57 to slide up and down in the rotating sleeve 58 in a reciprocating manner by being matched with the adjusting groove 541, and then the supporting plate 6 is driven to shake synchronously by the shaking rod 57 in the rotating process of the supporting plate 6, so that the culture solution in the culture dish 7 is shaken uniformly, the problem that the nutrient components in the culture solution are uneven is avoided, the cell proliferation effect of animal tissues is influenced, and the shaking rod 57 can be compensated by the upper moving connecting block 59 and the lower moving connecting block 59 in the horizontal direction.
When the culture solution in the culture dish 7 is insufficient, the sealing plug 87 is opened, the culture solution is filled into the upper charging barrel 82 through the feeding hole, then the sealing plug 87 is closed, according to the amount of the culture solution required in each culture dish 7, when the culture solution required in each culture dish 7 is the same, the valve 86 is synchronously opened, the air is extruded to the upper charging barrel 82 through the injection barrel 83, the extruded air is mutually matched with the connecting pipe 85 through the liquid distribution pipe 84 to pump the culture solution into the culture dish 7, when the culture solution required in each culture dish 7 is different, the valve 86 is respectively opened, the injection barrel 83 extrudes the air through the injection barrel 83, the culture solution is mutually matched with the connecting pipe 85 through the liquid distribution pipe 84 to pump the culture solution into the culture dish 7, the possibility that the mixed bacteria are introduced in the process of directly adding the culture solution into the culture dish 7 through manually opening the cover plate 4 is avoided, wherein the scales are marked on the injection barrel 83, and the culture solution can be quantitatively added into the culture dish 7, the feeding cylinder 82 and the connecting pipe 85 are detachably mounted on the feeding column 81 and the liquid separating pipe 84 respectively, so that the subsequent quick detachment and cleaning of the feeding cylinder 82 and the connecting pipe 85 are facilitated.
As shown in fig. 7, a rotating shaft 821 is installed in the middle of the lower end face of the interior of the upper charging barrel 82 through a bearing, a plurality of stirring claws 822 are evenly arranged below the circumferential face of the rotating shaft 821, a first bevel gear 823 is installed above the rotating shaft 821, a linkage shaft 824 is installed above the right side wall of the upper charging barrel 82 through a bearing, a second bevel gear 825 engaged with the first bevel gear 823 is installed on the left end face of the linkage shaft 824, a transmission gear 826 is installed on the linkage shaft 824, a sliding plate 827 is vertically penetrated and arranged at the upper end of the upper charging barrel 82, the upper end of the sliding plate 827 abuts against the lower end face of the telescopic end of the injection barrel 83, a rack matched with the transmission gear 826 is arranged at the lower end of the front end face of the sliding plate 827, a horizontal fixing protrusion is arranged at the bottom of the sliding plate 827, a return spring bar 828 is installed between the sliding plate 827 and the inner wall of the upper charging barrel 82 through the fixing protrusion, after the culture liquid is poured into the interior of the upper charging barrel 82, the culture liquid is in a standing state, the inside nutrient composition of culture solution can take place the phenomenon of sediment and lead to the inside inhomogeneous problem of nutrient substance that exists of culture solution, so when pressing into injection tube 83 and go into the culture solution to culture dish 7 in, sliding plate 827 moves down in step this moment, sliding plate 827 drives transmission gear 826 through the rack and rotates, and then transmission gear 826 drives universal driving shaft 824 and rotates, universal driving shaft 824 drives rotation axis 821 through a bevel gear 823 and No. two bevel gear 825 mutually supporting, thereby rotation axis 821 drives stirring claw 822 and homogenizes the processing to the culture solution, improve the inside nutrient substance's of culture solution homogeneity, wherein reset spring pole 828 plays the effect of resetting to sliding plate 827.
As shown in fig. 3 and 6, four the cavity inner wall and be located the bearing tray 6 outside and all install inside for loop forming element 61, and loop forming element 61 sets up the structure that the groove was led to the circle for square board up end, evenly install a plurality of bearing spring rods 62 on the loop forming element 61 inner wall, bearing spring rod 62 keeps away from loop forming element 61's one end and installs and bear tray 6 matched with bearing piece 63 through the ball pivot, loop forming element 61 can play spacing bearing effect to bearing tray 6 through bearing spring rod 62 and bearing piece 63 mutually supporting, avoid bearing tray 6 single-point atress instability on vertical even axle 56, increase bearing tray 6 and rock the in-process stable effect.
As shown in figure 8 of the drawings, as shown in fig. 9 and 10, a plurality of guide plates 851 provided with guide holes are uniformly arranged inside the connecting pipe 85, a drainage plate 852 is installed at the outlet of the liquid distribution pipe 84 on the lower end face of the cover plate 4 and below the lower end face of the cover plate through a support frame, the upper end face of the drainage plate 852 is set to be a conical structure, the lower end of the drainage plate 852 is set to be an umbrella-shaped structure, a plurality of drainage grooves 853 are uniformly arranged on the upper end face of the umbrella-shaped structure in the circumferential direction, the drainage plate 852 with the lower end being the umbrella-shaped structure can guide the culture solution through being matched with the drainage grooves 853, the problem that the distribution of the culture solution is not uniform when the culture solution enters the culture dish 7, the problem that the nutrient content of the animal tissue cells is not uniform is caused, the proliferation effect of the animal tissue cells is affected, the impact force when the culture solution enters the culture dish 7 can be reduced by the guide plates 851, and the problem that the animal tissue cells are scattered by the excessive impact force of the culture solution, and the culture condition of the animal tissue cells is affected.
As shown in fig. 9, connecting springs 854 are symmetrically installed between the guiding plate 851 and the inner wall of the connecting pipe 85 from top to bottom, wiping sponges 855 are disposed at the portions of the guiding plate 851, which are in contact with the inner wall of the connecting pipe 85, and guiding ropes 856 are installed between two adjacent guiding plates 851 located on the same side of the valve 86.
As shown in fig. 11, the present invention also provides a method for culturing animal cells, comprising the steps of: the first step is as follows: placing treatment: adding culture solution into the culture dish 7 filled with the animal tissue cells, taking down the cover plate 4, then clamping the culture dish 7 in the bearing tray 6, and finally closing the cover plate 4.
The second step is that: shaking treatment: the shaking motor 51 is started to drive the rotating shaft 52 to rotate through a gear meshing mode, the rotating shaft 52 drives the vertical moving block 55 to synchronously rotate through the rotating disc 53, the vertical moving block 55 drives the bearing plate 6 to rotate around the vertical connecting shaft 56 through the shaking rod 57, the vertical moving block 55 drives the shaking rod 57 to vertically slide in the rotating sleeve 58 through mutual matching with the adjusting groove 541 in the rotating process of the bearing plate 6, and then the bearing plate 6 is synchronously driven to shake through the shaking rod 57 in the rotating process of the bearing plate 6, so that the culture solution in the culture dish 7 is shaken uniformly.
The third step: and (3) loading treatment: when the culture solution in the culture dish 7 is not enough, open sealing plug 87 and go up the inside culture solution of pouring into of feed cylinder 82 through the feed inlet, close sealing plug 87 again, according to how much of required culture solution in each culture dish 7, open valve 86 and pass through injection tube 83 to feed cylinder 82 extrusion air, extruded air is mutually supported through branch liquid pipe 84 and connecting pipe 85 and is gone into the culture dish 7 with the culture solution pump, injection tube 83 presses in-process this moment sliding plate 827 and moves down in step, sliding plate 827 passes through the rack and drives transmission gear 826 and rotate, and then transmission gear 826 drives universal driving shaft 824 and rotates, universal driving shaft 824 drives the rotation shaft 821 through a bevel gear 823 and No. two bevel gear 825 mutually supporting, thereby rotation shaft 821 drives the stirring claw 822 and homogenizes the processing to the culture solution.
The fourth step: and (4) observation and taking out: observing the growth condition of the animal tissue cells through the microscopic magnifier 41, opening the cover plate 4 to take out the culture dish 7 after the culture of the animal tissue cells is finished, and finishing the work.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. The utility model provides an animal cell tissue culture device, includes culture platform (1), spliced pole (2), culture frame (3), apron (4), rocks mechanism (5), holds tray (6), culture dish (7) and feed mechanism (8), its characterized in that: cultivation platform (1) four corners of up end install spliced pole (2), culture frame (3) are installed jointly to four spliced pole (2) up end, culture frame (3) are inside to be separated into four cavitys through the cross baffle, four cavitys are inside and culture platform (1) up end is installed jointly and is rocked mechanism (5), rock mechanism (5) upper end and install bearing dish (6), the joint has culture dish (7) on bearing dish (6), apron (4) are installed to culture frame (3) up end, feed mechanism (8) are installed to apron (4) up end, apron (4) up end just is located the cavity and installs microscope magnifier (41) of observing the cavity directly over, observation window (42) have been seted up to four of culture frame (3) lateral wall, wherein:
the shaking mechanism (5) comprises a shaking motor (51), a rotating shaft (52), a rotating disc (53), an annular adjusting plate (54), a vertical moving block (55), a vertical connecting shaft (56), a shaking rod (57) and a rotating sleeve (58), wherein the shaking motor (51) is installed in the middle of the upper end face of the culture platform (1) through a motor base, the rotating shaft (52) is installed at the lower end of the culture frame (3) and positioned in the middle of four cavities in a penetrating manner through bearings, output shafts of the shaking motor (51) are connected with the four rotating shafts (52) in a gear meshing manner, the rotating disc (53) is installed on the upper end face of the rotating shaft (52), the annular adjusting plate (54) is installed in the middle of the bottom of the cavity, a plurality of adjusting grooves (541) are evenly formed in the circumferential direction of the upper end face of the annular adjusting plate (54), the vertical moving block (55) is arranged on the upper end of the rotating disc (53) in a penetrating and sliding manner up and down, the lower end of a vertical moving block (55) abuts against an annular adjusting plate (54), the middle of the upper end face of a rotating disc (53) is provided with a vertical connecting shaft (56) through a bearing, the upper end face of the vertical connecting shaft (56) is provided with a bearing tray (6) through a spherical hinge, the middle of the vertical connecting shaft (56) is provided with a through groove (561) from left to right, a rotating sleeve (58) is arranged in the through groove (561) through a hinge shaft, a shaking rod (57) is slidably arranged in the rotating sleeve (58), the lower end face of the bearing tray (6) and the upper end face of the vertical moving block (55) are both horizontally slidably provided with a moving connecting block (59), the upper end of the shaking rod (57) is arranged on the upper moving connecting block (59) through a hinge, and the lower end of the shaking rod (57) is arranged on the lower moving connecting block (59) through a hinge;
the feeding mechanism (8) comprises a feeding column (81), a feeding barrel (82), an injection cylinder (83), a liquid distribution pipe (84) and a connecting pipe (85), wherein material loading post (81) install in apron (4) up end middle part, it installs in material loading post (81) upper end to go up feed cylinder (82) through threaded connection mode, the feed inlet of installation sealing plug (87) is seted up on material loading cylinder (82) left side, it installs injection tube (83) that link up mutually and have the scale with last feed cylinder (82) to go up feed cylinder (82) up end, go up feed cylinder (82) lower terminal surface circumference and evenly install a plurality of liquid distribution pipes (84) that link up mutually with last feed cylinder (82), apron (4) up end just is located also to install liquid distribution pipe (84) that link up mutually with the cavity directly over four culture dishes (7), the joint has connecting pipe (85) that have valve (86) between two upper and lower adjacent liquid distribution pipes (84).
2. An animal cell tissue culture apparatus according to claim 1, wherein: go up inside lower terminal surface middle part of feed cylinder (82) install axis of rotation (821) through the bearing, axis of rotation (821) periphery below evenly is provided with a plurality of stirring claw (822), axis of rotation (821) top is installed bevel gear (823), go up feed cylinder (82) right side wall top and install universal driving shaft (824) through the bearing, universal driving shaft (824) left end face is installed and is had No. two bevel gear (825) of bevel gear (823) engaged with, install drive gear (826) on universal driving shaft (824), it is provided with sliding plate (827) to run through from top to bottom to go up feed cylinder (82) upper end, sliding plate (827) upper end supports and leans on at injection tube (83) flexible end lower extreme, sliding plate (827) front end face lower extreme is provided with drive gear (826) matched with rack, sliding plate (827) bottom is provided with horizontal fixed arch, install reset spring pole (828) between fixed arch and last feed cylinder (82) inner wall sliding plate (827).
3. An animal cell tissue culture apparatus according to claim 1, wherein: four cavity inner wall and be located and all install loop forming element (61) in bearing tray (6) outside, and loop forming element (61) are square board up end and set up the structure that the groove was led to the circle, evenly install a plurality of bearing spring levers (62) on loop forming element (61) inner wall, bearing spring lever (62) keep away from the one end of loop forming element (61) and install and bear tray (6) matched with bearing piece (63) through the ball pivot.
4. An animal cell tissue culture apparatus according to claim 1, wherein: connecting pipe (85) inside evenly set up a plurality of guide plates (851) of seting up the guide hole, drainage plate (852) are installed through the support frame in liquid distribution pipe (84) exit that terminal surface just is located the below under apron (4), drainage plate (852) up end sets up to the toper structure, drainage plate (852) lower extreme sets up to the umbrella type structure, umbrella type structure up end circumference evenly is provided with a plurality of drainage grooves (853).
5. An animal cell tissue culture apparatus according to claim 4, wherein: connecting springs (854) are symmetrically arranged between the guide plates (851) and the inner wall of the connecting pipe (85) from top to bottom, wiping sponges (855) are arranged on the contact part of the guide plates (851) and the inner wall of the connecting pipe (85), and guide ropes (856) are arranged between two adjacent guide plates (851) on the same side of the valve (86).
6. An animal cell tissue culture apparatus according to claim 1, wherein: the edge of the lower end face of the cover plate (4) is provided with a rectangular protrusion (32), and the edge of the upper end face of the culture frame (3) is provided with a sealing groove (31) matched with the rectangular protrusion (32).
7. An animal cell tissue culture apparatus according to claim 1, wherein: the inner wall of the culture dish (7) is clamped with an annular plate (71).
8. An animal cell tissue culture apparatus according to claim 1, wherein: the depths of the adjacent adjusting grooves (541) are different.
9. An animal cell tissue culture apparatus according to claim 1, wherein: the method for culturing the animal cells by using the animal cell tissue culture device comprises the following steps:
the first step is as follows: placing treatment: adding nutrient solution into the culture dish (7) filled with the animal tissue cells, taking down the cover plate (4), clamping the culture dish (7) in the bearing tray (6), and finally closing the cover plate (4);
the second step is that: shaking treatment: the shaking motor (51) is started to drive the rotating shaft (52) to rotate in a gear transmission mode, the rotating shaft (52) drives the vertical moving block (55) to reciprocate up and down on the annular adjusting plate (54) through the rotating disc (53), and then the vertical moving block (55) drives the bearing plate (6) to shake synchronously in the rotating process of the bearing plate (6) through the shaking rod (57), so that the nutrient solution in the culture dish (7) is shaken, shaken and uniformly shaken;
the third step: and (3) loading treatment: when the nutrient solution in the culture dish (7) is insufficient, opening a sealing plug (87) to fill the nutrient solution into the upper charging barrel (82) through a feeding hole, closing the sealing plug (87), opening a valve (86) to extrude air through an injection tube (83) according to the amount of the required nutrient solution in each culture dish (7), and mutually matching the nutrient solution with a connecting tube (85) through a liquid distribution tube (84) to add the nutrient solution into the culture dish (7);
the fourth step: and (4) observation and taking out: and (3) observing the growth condition of the animal tissue cells through a microscopic magnifier (41), opening the cover plate (4) to take out the culture dish (7) after the culture of the animal tissue cells is finished, and finishing the work.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210033187.XA CN114369533A (en) | 2022-01-12 | 2022-01-12 | Animal cell tissue culture device and culture method |
| PCT/CN2022/144047 WO2023134476A1 (en) | 2022-01-12 | 2022-12-30 | Animal cell tissue culture device and culture method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210033187.XA CN114369533A (en) | 2022-01-12 | 2022-01-12 | Animal cell tissue culture device and culture method |
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| CN114369533A true CN114369533A (en) | 2022-04-19 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202210033187.XA Withdrawn CN114369533A (en) | 2022-01-12 | 2022-01-12 | Animal cell tissue culture device and culture method |
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| Country | Link |
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| CN (1) | CN114369533A (en) |
| WO (1) | WO2023134476A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115747059A (en) * | 2022-11-30 | 2023-03-07 | 湖南臻和亦康医疗用品有限公司 | Cell tissue culture environment simulation device |
| WO2023134476A1 (en) * | 2022-01-12 | 2023-07-20 | 阜阳师范大学 | Animal cell tissue culture device and culture method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110684660A (en) * | 2019-11-01 | 2020-01-14 | 力盟生命科技(深圳)有限公司 | Micro-vibration culture device for embryos |
| CN115926956A (en) * | 2023-02-07 | 2023-04-07 | 山东灵犀生物科技有限公司 | Cell culture exosome collection device |
| CN117343837B (en) * | 2023-10-07 | 2024-04-05 | 中国人民解放军总医院第六医学中心 | Stem cell continuous liquid-changing culture device and application method thereof |
| CN117106555B (en) * | 2023-10-25 | 2024-03-01 | 山东启腾生物科技有限公司 | Bean product fermented milk fungus bacterial culture apparatus |
| CN117683634B (en) * | 2024-01-31 | 2024-05-07 | 山东格林医学科技有限公司 | Stem cell culture device and method for diabetes treatment |
| CN117683613B (en) * | 2024-02-04 | 2024-04-19 | 科泰(山东)健康营养有限公司 | Experimental equipment for influence of choline bitartrate on lung cancer cells |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109852546B (en) * | 2019-04-03 | 2020-01-31 | 崔建英 | Pollution-free culture system and method for cells |
| CN112342122A (en) * | 2020-11-11 | 2021-02-09 | 何伟 | Anti-pollution type bacterial culture device and bacterial culture method for clinical laboratory |
| CN113755329A (en) * | 2021-09-23 | 2021-12-07 | 赵宝玉 | Cell culture dish |
| CN113832009A (en) * | 2021-09-24 | 2021-12-24 | 曾峰 | A rotatable formula cell culture dish for cell culture |
| CN114369533A (en) * | 2022-01-12 | 2022-04-19 | 阜阳师范大学 | Animal cell tissue culture device and culture method |
-
2022
- 2022-01-12 CN CN202210033187.XA patent/CN114369533A/en not_active Withdrawn
- 2022-12-30 WO PCT/CN2022/144047 patent/WO2023134476A1/en unknown
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023134476A1 (en) * | 2022-01-12 | 2023-07-20 | 阜阳师范大学 | Animal cell tissue culture device and culture method |
| CN115747059A (en) * | 2022-11-30 | 2023-03-07 | 湖南臻和亦康医疗用品有限公司 | Cell tissue culture environment simulation device |
| CN115747059B (en) * | 2022-11-30 | 2024-03-29 | 湖南臻和亦康医疗用品有限公司 | Cell tissue culture environment simulation device |
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| WO2023134476A1 (en) | 2023-07-20 |
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