WO2023134476A1 - 一种动物细胞组织培养装置及培养方法 - Google Patents

一种动物细胞组织培养装置及培养方法 Download PDF

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Publication number
WO2023134476A1
WO2023134476A1 PCT/CN2022/144047 CN2022144047W WO2023134476A1 WO 2023134476 A1 WO2023134476 A1 WO 2023134476A1 CN 2022144047 W CN2022144047 W CN 2022144047W WO 2023134476 A1 WO2023134476 A1 WO 2023134476A1
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culture
end surface
plate
shaking
cover plate
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PCT/CN2022/144047
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English (en)
French (fr)
Inventor
姬云涛
夏珍妮
王硕
陈清怡
蒋侠森
屈长青
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阜阳师范大学
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Publication of WO2023134476A1 publication Critical patent/WO2023134476A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/38—Caps; Covers; Plugs; Pouring means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/10—Petri dish
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/22—Transparent or translucent parts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M27/16—Vibrating; Shaking; Tilting
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M27/18—Flow directing inserts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/10—Perfusion
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor

Definitions

  • 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.
  • Animal cell culture is a method of taking out relevant tissues from the animal body, placing the tissues in the culture medium to allow these tissue cells to grow and proliferate to obtain new cells.
  • cell culture technology has become It has become the basis of transgenic technology, biopharmaceuticals and many other technologies, and plays an important role in modern biotechnology.
  • the traditional culture medium replenishment process needs to manually open the culture device and directly add the culture medium to the culture dish. This process may cause airborne bacteria to enter the culture medium. In the dish, miscellaneous bacteria enter the culture solution and affect the growth of animal tissue cells, and even harmful miscellaneous bacteria may cause the proliferation of animal tissue cells to stop, affecting the results of animal tissue cell culture.
  • a kind of animal cell tissue culture device including culture platform, connecting column, culture frame, cover plate, shaking mechanism, supporting tray, culture dish and feeding mechanism, described culture Connecting columns are installed at the four corners of the upper end of the platform, and the upper ends of the four connecting columns are jointly installed with a culture frame.
  • the inside of the culture frame is divided into four cavities by cross-shaped partitions.
  • a shaking mechanism is installed, and a supporting tray is installed on the upper end of the shaking mechanism, and a culture dish is clamped on the supporting tray.
  • a microscope magnifying glass for the observation cavity is installed directly above, and observation windows are opened at the four corners of the side wall of the culture frame.
  • the shaking mechanism includes a shaking motor, a rotating shaft, a rotating disc, an annular adjustment plate, a vertical moving block, a vertical connecting shaft, a shaking rod and a rotating sleeve, wherein the shaking motor is installed on the cultivation platform through a motor base In the middle of the end face, the lower end of the cultivation frame is located in the middle of the four chambers, and a rotating shaft is installed through bearings.
  • the output shaft of the shaking motor and the four rotating shafts are connected to each other through gear meshing.
  • a rotating disc is installed on the upper end of the rotating shaft, and the bottom of the chamber
  • a ring-shaped adjustment plate is installed in the middle position, and the upper surface of the ring-shaped adjustment plate is evenly provided with a plurality of adjustment grooves in the circumferential direction.
  • the upper end of the rotating disc slides through and is provided with a vertical moving block, and the lower end of the vertical moving block is against the ring-shaped adjusting plate.
  • the middle part of the upper end surface of the rotating disk is installed with a vertical shaft through a bearing, and the upper end of the vertical shaft is equipped with a bearing tray through a ball hinge.
  • the shaft is equipped with a rotating sleeve, and the inside of the rotating sleeve is slidingly provided with a rocking rod.
  • the lower end surface of the supporting tray and the upper end face of the vertical moving block are both horizontally slidingly provided with a moving connecting block, and the upper end of the rocking rod is installed on the upper moving connecting block through a hinge.
  • the lower end of the rocking rod is installed on the moving connecting block below through a hinge.
  • the feeding mechanism includes a feeding column, a feeding cylinder, an injection cylinder, a liquid pipe and a connecting pipe, wherein the feeding column is installed in the middle of the end surface of the cover plate, and the feeding cylinder is installed on the upper surface through a threaded connection.
  • On the upper surface of the upper material cylinder there is an injection cylinder connected with the upper material cylinder and with a scale. The lower end surface of the upper material cylinder is evenly installed in the circumferential direction.
  • the middle part of the inner lower end surface of the upper material cylinder is equipped with a rotating shaft through a bearing, a plurality of stirring claws are uniformly arranged under the circumference of the rotating shaft, a No.
  • the bearing is equipped with a linkage shaft, and the left end of the linkage shaft is equipped with a No. 2 bevel gear that meshes with the No. 1 bevel gear.
  • the transmission gear is installed on the linkage shaft.
  • a sliding plate is installed up and down on the upper end of the upper barrel, and the upper end of the sliding plate is against the
  • the lower end of the telescopic end of the injection barrel, the lower end of the front end of the sliding plate is provided with a rack that matches the transmission gear, the bottom of the sliding plate is provided with a horizontal fixing protrusion, and the sliding plate is installed between the fixing protrusion and the inner wall of the upper barrel. spring bar.
  • the inner walls of the four cavities and the outer sides of the supporting tray are all equipped with ring pieces
  • the ring piece is a structure in which a round through groove is opened on the end surface of the square plate, and a plurality of supporting spring rods are uniformly installed on the inner wall of the ring piece
  • the end of the supporting spring rod far away from the ring part is equipped with a supporting block that matches the supporting tray through a ball joint
  • the ring part cooperates with the supporting spring rod and the supporting block to limit the supporting effect on the supporting tray, avoiding
  • the single-point force instability of the support tray on the vertical shaft increases the stability of the support tray during the shaking process.
  • a plurality of guide plates with guide holes are evenly arranged inside the connecting pipe, and a drain plate is installed on the lower end surface of the cover plate and at the outlet of the dispensing pipe below through a support frame, and the upper surface of the drain plate is set as Conical structure, the lower end of the drainage plate is set as an umbrella structure, and a plurality of drainage grooves are uniformly arranged on the upper surface of the umbrella structure, and the lower end of the drainage plate with an umbrella structure can guide the culture medium by cooperating with the drainage grooves , to avoid the uneven distribution of the culture solution when it enters the culture dish, which will cause the problem that the animal tissue cells obtain uneven nutrients, and affect the proliferation effect of the animal tissue cells.
  • the guide plate can reduce the impact force of the culture solution entering the culture dish, and avoid The excessive impact force of the culture medium disperses the animal tissue cells, which affects the culture of the animal tissue cells.
  • connecting springs are installed symmetrically up and down between the guide plate and the inner wall of the connecting pipe, and the part of the guide plate in contact with the inner wall of the connecting pipe is provided with a wiping sponge, which is located between two adjacent guide plates on the same side of the valve.
  • a guide rope is installed. Since the diameter of the connecting pipe is small and long, the guide rope is manually driven to drive the guide plate to slide back and forth inside the connecting pipe during the cleaning process of the connection pipe, and then the guide plate can be cleaned by wiping the sponge.
  • the inner wall of the connecting pipe is reciprocally wiped and cleaned, which is convenient for cleaning the inside of the connecting pipe, wherein the connecting spring resets the guide plate during the cleaning process.
  • the edge of the lower end surface of the cover plate is provided with a rectangular protrusion
  • the edge of the upper end surface of the cultivation frame is provided with a sealing groove matching with the rectangular protrusion, and the sealing groove and the rectangular protrusion can cooperate with each other to seal the inside of the cavity. , to prevent impurities in the air from entering the cavity and affecting the growth of animal tissue cells.
  • annular plate is clamped on the inner wall of the petri dish, and the annular plate is snapped on the inner wall of the petri dish after the addition of the animal tissue cells and culture solution in the petri dish is completed, so as to prevent the culture liquid from flowing out of the petri dish during the sliding process of the petri dish.
  • Splashing in the medium causes waste of the culture medium, and at the same time ensures the environmental sanitation inside the cavity.
  • the depths of the adjacent adjustment grooves are different, so that the vertical moving block obtains different amplitudes in the process of moving up and down, and then drives the supporting tray to obtain different shaking amplitudes through the shaking rod, thereby ensuring that the culture dish obtains different shaking amplitudes so that The distribution of nutrients in the culture medium is more uniform.
  • the method for cultivating animal cells by the animal cell tissue culture device includes the following steps: the first step: placing treatment: adding culture solution to the inside of the petri dish containing animal tissue cells, removing the cover plate and then Snap the Petri dish inside the tray, and finally close the cover.
  • Step 2 Shaking treatment: the shaking motor starts to drive the rotating shaft to rotate through the gear transmission mode, and the rotating shaft drives the vertical moving block to move up and down on the annular adjustment plate by rotating the disc, and then the vertical moving block drives the bearing tray to rotate through the shaking rod Synchronously drives the supporting tray to shake, so as to shake and shake the culture solution in the culture dish evenly.
  • Step 3 Feeding treatment: When the culture medium in the culture dish is insufficient, open the sealing plug and pour the culture liquid into the upper cylinder through the feeding port, and then close the sealing plug, according to the culture liquid required in each culture dish How much, open the valve and squeeze the air through the syringe to transfer the culture solution through the dispensing tube and the connecting tube to cooperate with each other to add the culture solution into the culture dish.
  • Step 4 Observation and taking out: Observe the growth of animal tissue cells through a microscope magnifying glass. When the animal tissue cells are cultured, open the cover and take out the culture dish, and the work is over.
  • the beneficial effects of the present invention are: 1.
  • the culture frame designed by the present invention is divided into four cavities, which can simultaneously proliferate and cultivate the animal cell tissues in the four culture dishes during the subculture of animal cell tissues, thereby improving the quality of animal cell tissues. Proliferation and cultivation efficiency; in the rocking mechanism, the rotating disc drives the vertical moving block to move up and down on the circular adjustment plate, and then the vertical moving block drives the supporting tray to shake synchronously during the rotation process through the shaking rod, so that the culture dish Shake the culture solution in the feeding mechanism; in the feeding mechanism, open the sealing plug and pour the culture solution into the upper barrel through the feeding port, and then close the sealing plug, according to the amount of culture solution required in each culture dish , open the valve and squeeze the air through the injection barrel to pump the culture solution into the culture dish through the cooperation of the dispensing tube and the connecting tube.
  • the cover plate is not opened during the whole feeding process, thereby reducing the possibility of bacteria entering the cavity Possibly, reduce the possibility of bacteria contaminating animal tissue cell culture.
  • 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 diagram of the internal three-dimensional structure of the culture frame of the present invention.
  • Fig. 4 is a schematic diagram of the internal installation structure of the cavity and the shaking mechanism of the present invention.
  • Fig. 5 is a partial sectional view at A of Fig. 4 of the present invention.
  • Fig. 6 is a schematic diagram of the three-dimensional installation structure between the cavity and the shaking mechanism of the present invention.
  • Fig. 7 is a schematic diagram of the internal structure of the upper barrel of the present invention.
  • Fig. 8 is a schematic diagram of the internal structure of the connecting pipe of the present invention.
  • Fig. 9 is a partial enlarged view of B in Fig. 8 of the present invention.
  • Fig. 10 is a schematic diagram of the three-dimensional structure of the drainage plate of the present invention.
  • Fig. 11 is a flowchart of animal cell tissue culture in the present invention.
  • an animal cell tissue culture device includes a culture platform 1, a connecting column 2, a culture frame 3, a cover plate 4, a shaking mechanism 5, a supporting tray 6, a culture dish 7 and an upper Material mechanism 8, the four corners of the upper end surface of the cultivation platform 1 are equipped with connecting columns 2, and the upper end surfaces of the four connecting columns 2 are jointly equipped with a cultivation frame 3, and the inside of the cultivation frame 3 is divided into four chambers by cross-shaped partitions Body, inside the four cavities and the upper end of the cultivation platform 1 are installed with a shaking mechanism 5, the upper end of the shaking mechanism 5 is installed with a supporting tray 6, the supporting tray 6 is clamped with a culture dish 7, and the upper end of the cultivation frame 3 is equipped with a cover plate 4.
  • a feeding mechanism 8 is installed on the upper end of the cover plate 4, and a microscopic magnifying glass 41 for observing the cavity is installed on the upper end surface of the cover plate 4 and directly above the cavity, and observation windows are provided at the four corners of the side walls of the cultivation frame 3 42.
  • the edge of the lower end surface of the cover plate 4 is provided with a rectangular protrusion 32
  • the edge of the upper end surface of the cultivation frame 3 is provided with a sealing groove 31 that matches the rectangular protrusion 32, and the sealing groove 31 is in contact with the rectangular protrusion 32.
  • the mutual cooperation of the rectangular protrusions 32 can seal the interior of the cavity, preventing impurities in the air from entering the cavity and affecting the growth of animal tissue cells.
  • annular plate 71 is clamped on the inner wall of the petri dish 7.
  • the annular plate 71 is clamped on the inner wall of the petri dish 7 to prevent the 7 .
  • the culture solution splashes out from the culture dish 7, causing waste of the culture solution, while ensuring the sanitation inside the cavity.
  • the shaking mechanism 5 includes a shaking motor 51, a rotating shaft 52, a rotating disk 53, an annular adjustment plate 54, a vertical moving block 55, and a vertical connecting shaft 56. , a rocking rod 57 and a rotating sleeve 58, wherein the rocking motor 51 is installed on the middle part of the upper end surface of the cultivation platform 1 through the motor base, and the lower end of the culture frame 3 is installed in the middle of the four cavities through bearings. 51 The output shaft and the four rotating shafts 52 are all connected to each other through gear meshing. A rotating disk 53 is installed on the upper end of the rotating shaft 52. An annular adjustment plate 54 is installed at the middle of the bottom of the cavity.
  • the upper end of the annular adjustment plate 54 is uniform in the circumferential direction.
  • a plurality of adjustment grooves 541 are provided, and the upper end of the rotating disc 53 slides up and down to be provided with a vertical moving block 55, and the lower end of the vertical moving block 55 is against the annular adjusting plate 54, and the middle part of the upper end surface of the rotating disc 53 passes through the bearing
  • a vertical connecting shaft 56 is installed, and the upper end surface of the vertical connecting shaft 56 is equipped with a bearing tray 6 through a ball hinge.
  • the middle part of the vertical connecting shaft 56 is provided with a through groove 561 from left to right, and the inside of the through groove 561 is equipped with a rotating shaft through a hinge shaft.
  • Sleeve 58, rotating sleeve 58 internal sliding is provided with rocking bar 57, and the lower end surface of bearing tray 6 and the upper end surface of vertical moving block 55 are all horizontally slidingly provided with moving connecting block 59, and the upper end of rocking bar 57 is installed on the moving side by hinge.
  • the connecting block 59, the lower end of the rocking bar 57 is installed on the moving connecting block 59 below by a hinge.
  • the depths of the adjacent adjustment grooves 541 are different, so that the vertical moving block 55 obtains different amplitudes during the up and down movement process, and then drives the supporting tray 6 through the rocking rod 57 to obtain different shaking amplitudes , so as to ensure that the petri dish 7 obtains different shaking ranges so that the nutrient components in the culture solution are distributed more evenly.
  • the feeding mechanism 8 includes a feeding column 81, a feeding cylinder 82, a syringe 83, a liquid pipe 84 and a connecting pipe 85, wherein the loading column 81 is installed on the cover plate 4 In the middle part of the upper end face, the upper material cylinder 82 is installed on the upper end of the upper material column 81 through a threaded connection, and the left side of the upper material cylinder 82 is provided with a feed port for installing a sealing plug 87.
  • the injection cylinder 83 is connected and has a scale.
  • the lower end surface of the upper cylinder 82 is evenly installed with a plurality of liquid pipes 84 connected with the upper cylinder 82 in the circumferential direction.
  • the upper end surface of the cover plate 4 is located on the front of the four culture dishes 7
  • a dispensing pipe 84 connected to the cavity is installed on the top, and a connecting pipe 85 with a valve 86 is clamped between the upper and lower adjacent dispensing pipes 84 .
  • the block 55 cooperates with the adjustment groove 541 to drive the shaking rod 57 to reciprocate and slide up and down inside the rotating sleeve 58, and then drives the supporting tray 6 to shake synchronously through the shaking rod 57 during the rotation process of the supporting tray 6, so that the culture dish 7 Shake the culture solution evenly to avoid uneven nutrition in the culture solution and affect the proliferation of animal tissue cells.
  • the upper and lower moving connecting blocks 59 can compensate for the displacement of the shaking rod 57 in the horizontal direction.
  • the middle part of the inner lower end surface of the upper material cylinder 82 is equipped with a rotating shaft 821 through a bearing, a plurality of stirring claws 822 are evenly arranged under the circumference of the rotating shaft 821, and a No. 1 bevel gear 823 is installed above the rotating shaft 821
  • a linkage shaft 824 is mounted on the top of the right side wall of the upper material cylinder 82 through a bearing, and the second bevel gear 825 meshing with the first bevel gear 823 is installed on the left end of the linkage shaft 824.
  • the upper end of the barrel 82 is provided with a sliding plate 827 up and down. The upper end of the sliding plate 827 is against the lower end surface of the telescopic end of the injection barrel 83.
  • the lower end of the front end of the sliding plate 827 is provided with a rack that matches the transmission gear 826.
  • the sliding plate 827 is provided with a return spring rod 828 between the fixed protrusions and the inner wall of the upper material cylinder 82.
  • the linkage shaft 824 drives the rotation shaft 821 to rotate through the cooperation of the No. 1 bevel gear 823 and the No. 2 bevel gear 825, so that the rotation shaft 821 drives the stirring claw 822 stirs the culture solution to improve the uniformity of nutrients inside the culture solution, wherein the return spring rod 828 resets the sliding plate 827.
  • the inner walls of the four chambers and the outer sides of the supporting tray 6 are all equipped with a ring part 61 inside, and the ring part 61 is a structure in which the upper surface of a square plate is provided with a round through groove.
  • a plurality of supporting spring rods 62 are evenly installed on the inner wall of 61, and one end of the supporting spring rods 62 away from the ring part 61 is equipped with a supporting block 63 matched with the supporting tray 6 through a ball hinge, and the ring part 61 passes through the supporting spring rods.
  • 62 cooperates with supporting block 63 and can play a position-limiting supporting effect to supporting tray 6, avoid single-point force instability of supporting tray 6 on vertical connecting shaft 56, and increase supporting tray 6 stabilizing effect in rocking process.
  • a plurality of guide plates 851 with guide holes are evenly arranged inside the connecting pipe 85, and the lower end surface of the cover plate 4 and the outlet of the liquid pipe 84 located below pass through
  • the support frame is equipped with a drainage plate 852, the upper end surface of the drainage plate 852 is set as a conical structure, the lower end of the drainage plate 852 is set as an umbrella structure, and a plurality of drainage grooves 853 are evenly arranged on the upper end surface of the umbrella structure, and the lower end is an umbrella structure.
  • the drainage plate 852 can guide the culture fluid by cooperating with the drainage groove 853, avoiding the uneven distribution of the culture fluid when it enters the culture dish 7, causing the problem that the animal tissue cells obtain uneven nutrients, and affecting the animal tissue cells.
  • the proliferating effect, the guide plate 851 can reduce the impact force of the culture solution entering the culture dish 7, avoiding the excessive impact force of the culture solution to disperse the animal tissue cells and affect the culture of the animal tissue cells.
  • connecting springs 854 are installed symmetrically up and down between the guide plate 851 and the inner wall of the connecting pipe 85 , and the part where the guide plate 851 contacts the inner wall of the connecting pipe 85 is provided with a wiping sponge 855 .
  • a guide rope 856 is installed between two adjacent guide plates 851 on the side. Since the diameter of the connecting pipe 85 is relatively small and long, the outermost two ends of the connecting pipe 85 are manually pulled when the connecting pipe 85 is passed through water for cleaning.
  • the first guide plate 851 moves, and then the two guide plates 851 drive the remaining guide plate 851 to slide back and forth inside the connecting pipe 85 through the guide rope 856, so that the guide plate 851 can clean the inner wall of the connecting pipe 85 by wiping the sponge 855 Reciprocating wiping and cleaning is convenient for cleaning the inside of the connecting pipe 85, wherein the connecting spring 854 resets the guide plate 851 during the cleaning process.
  • the present invention also provides a kind of culture method of animal cell, comprises the following steps: the first step: placement processing: add culture solution to the petri dish 7 inside that animal tissue cells are housed, remove cover plate 4 Then the petri dish 7 is clamped inside the supporting tray 6, and finally the cover plate 4 is closed.
  • the second step shaking processing: the shaking motor 51 starts to drive the rotating shaft 52 to rotate through the gear meshing mode, and the rotating shaft 52 drives the vertical moving block 55 to rotate synchronously through the rotating disc 53, and the vertical moving block 55 drives the supporting tray 6 to rotate through the shaking rod 57
  • the vertical moving block 55 cooperates with the adjustment groove 541 to drive the shaking rod 57 to reciprocate and slide up and down inside the rotating sleeve 58 during the rotating process of the supporting tray 6, and then passes through the rotating process of the supporting tray 6.
  • the shaking rod 57 synchronously drives the supporting tray 6 to shake, thereby shaking and shaking the culture solution in the culture dish 7 evenly.
  • the third step feeding process: when the nutrient solution in the petri dish 7 is not enough, open the sealing plug 87 and pour the nutrient solution into the upper barrel 82 through the feeding port, then close the sealing plug 87, according to each culture dish 7 How much culture solution is needed in the medium, open the valve 86 and squeeze the air through the injection cylinder 83 to the upper barrel 82, and the extruded air will pump the culture solution into the culture dish 7 through the liquid distribution pipe 84 and the connecting pipe 85, and the injection cylinder 83
  • the sliding plate 827 moves down synchronously at this time, the sliding plate 827 drives the transmission gear 826 to rotate through the rack, and then the transmission gear 826 drives the linkage shaft 824 to rotate, and the linkage shaft 824 passes through the No. 1 bevel gear 823 and the No. 2 bevel gear 825 Cooperate with each other to drive the rotating shaft 821 to rotate, so that the rotating shaft 821 drives the stirring claw 822 to stir the culture solution evenly.
  • Step 4 Observing and taking out: observe the growth of animal tissue cells through the magnifying glass 41, open the cover plate 4 to take out the culture dish 7 after the animal tissue cell culture is completed, and the work is finished.

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Abstract

本申请提供一种动物细胞组织培养装置及培养方法，涉及动物细胞组织培养的技术领域，该装置包括培养平台、连接柱、培养框、盖板、晃动机构、承托盘、培养皿以及上料机构，在晃动机构中，转动圆盘带动竖直移动块在环形调节板上上下往复移动，进而竖直移动块通过晃动杆带动承托盘转动过程中同步带动承托盘晃动，从而对培养皿中的营养液进行晃动摇匀处理；在上料机构中根据各个培养皿中所需要营养液的多少，打开阀门通过注射筒挤压空气将营养液通过分液管与连接管相互配合将营养液泵入培养皿中，在整个上料过程中未打开盖板，从而减少杂菌进入腔体内部的可能，减少杂菌污染动物组织细胞培养的可能性。

Description

一种动物细胞组织培养装置及培养方法 技术领域
本发明涉及动物细胞组织培养的技术领域，特别涉及一种动物细胞组织培养装置及培养方法。
 背景技术
动物细胞培养就是从动物体中取出相关的组织、将组织其放在培养液中让这些组织细胞生长和增殖得到新细胞的方法，随着基因工程和其他细胞工程技术的发展，细胞培养技术已成为转基因技术、生物制药及其他许多技术的基础，在现代生物技术中发挥着重要的作用。
人工通过刮板将刮下的动物组织细胞放置在培养皿中部，由于动物组织细胞在增殖生长过程中需要消耗培养液中的营养成分，故培养液中营养成分浓度在动物组织细胞培养过程中自培养皿中部向培养皿四周逐渐减小，培养液内部营养成分存在不均匀的问题，从而影响动物组织细胞增殖效果。
由于动物组织细胞在培养过程中可能需要及时补充培养液，传统的培养液补充过程中需要人工打开培养装置直接向培养皿中加入培养液，这个过程中可能会造成空气中的杂菌进入到培养皿中，导致杂菌进入培养液影响动物组织细胞生长，甚至有害杂菌可能会造成动物组织细胞增殖停止，影响动物组织细胞培养结果。
 技术问题
为了解决上述问题，本发明采用以下技术方案：一种动物细胞组织培养装置，包括培养平台、连接柱、培养框、盖板、晃动机构、承托盘、培养皿以及上料机构，所述的培养平台上端面四个拐角处安装有连接柱，四个连接柱上端面共同安装有培养框，培养框内部通过十字形隔板分隔成四个腔体，四个腔体内部以及培养平台上端面共同安装有晃动机构，晃动机构上端安装有承托盘，承托盘上卡接有培养皿，培养框上端面安装有盖板，盖板上端面安装有上料机构，盖板上端面且位于腔体的正上方安装有观测腔体的显微放大镜，培养框侧壁四个拐角处开设有观察窗。
所述的晃动机构包括晃动电机、转轴、转动圆盘、环形调节板、竖直移动块、竖直连轴、晃动杆以及转动套筒，其中所述的晃动电机通过电机座安装在培养平台上端面中部，培养框下端且位于四个腔体中部通过轴承贯穿安装有转轴，晃动电机输出轴与四个转轴之间均通过齿轮啮合方式相互连接，转轴上端面安装有转动圆盘，腔体底部中间位置安装有环形调节板，环形调节板上端面周向均匀开设有多个调节凹槽，转动圆盘上端上下贯穿滑动设置有竖直移动块，且竖直移动块下端抵靠在环形调节板上，转动圆盘上端面中部通过轴承安装有竖直连轴，竖直连轴上端面通过球铰安装有承托盘，竖直连轴中部自左向右开设有贯通槽，贯通槽内部通过铰轴安装有转动套筒，转动套筒内部滑动设置有晃动杆，承托盘下端面以及竖直移动块上端面均水平滑动设置有移动连块，晃动杆上端通过铰链安装在上方的移动连块上，晃动杆下端通过铰链安装在下方的移动连块上。
所述的上料机构包括上料柱、上料筒、注射筒、分液管以及连接管，其中所述的上料柱安装在盖板上端面中部，上料筒通过螺纹连接方式安装在上料柱上端，上料筒左侧开设有安装有密封塞的进料口，上料筒上端面安装有与上料筒相贯通且带有刻度的注射筒，上料筒下端面周向均匀安装有多个与上料筒相贯通的分液管，盖板上端面且位于四个培养皿正上方也安装有与腔体相贯通的分液管，上下两个相邻分液管之间卡接有带有阀门的连接管。
优选的，所述的上料筒内部下端面中部通过轴承安装有转动轴，转动轴圆周面下方均匀设置有多个搅拌爪，转动轴上方安装有一号锥齿轮，上料筒右侧壁上方通过轴承安装有联动轴，联动轴左端面安装有与一号锥齿轮相啮合的二号锥齿轮，联动轴上安装有传动齿轮，上料筒上端上下贯穿设置有滑动板，滑动板上端抵靠在注射筒伸缩端下端面，滑动板前端面下端设置有与传动齿轮相配合的齿条，滑动板底部设置有水平的固定凸起，滑动板通过固定凸起与上料筒内壁之间安装有复位弹簧杆。
优选的，四个所述的腔体内壁且位于承托盘外侧均安装有环形件，且环形件为方形板上端面开设有圆通槽的结构，环形件内壁上均匀安装有多个承托弹簧杆，承托弹簧杆远离环形件的一端通过球铰安装有与承托盘相配合的承托块，环形件通过承托弹簧杆与承托块相互配合可以对承托盘起限位承托作用，避免承托盘在竖直连轴上单点受力不稳定性，增加承托盘在晃动过程中稳定效果。
优选的，所述的连接管内部均匀设置多个开设有导引孔的导引板，盖板下端面且位于下方的分液管出口处通过支撑架安装有引流板，引流板上端面设置为锥形结构，引流板下端设置为伞型结构，伞型结构上端面周向均匀设置有多个引流槽，下端为伞形结构的引流板通过与引流槽相互配合可以对培养液起导流作用，避免培养液在进入培养皿时分布不均匀，造成动物组织细胞获得营养成分不均匀的问题，影响动物组织细胞的增殖效果，导引板可以减小培养液进入培养皿中的冲击力，避免培养液过大的冲击力冲散动物组织细胞，影响动物组织细胞的培养情况。
优选的，所述的导引板与连接管内壁之间上下对称安装有连接弹簧，导引板与连接管内壁接触的部分设置有擦拭海绵，位于阀门同一侧相邻两个导引板之间安装有导引绳，由于连接管直径较小且较长，故在对连接管进水清洗过程中手动带动导引绳带动导引板在连接管内部往复滑动，进而导引板通过擦拭海绵可以对连接管内壁进行往复擦拭清洗，便于对连接管内部的清理，其中连接弹簧在清洗过程中对导引板起复位作用。
优选的，所述的盖板下端面边缘处设置有矩形凸起，培养框上端面边缘开设有与矩形凸起相配合的密封槽，密封槽与矩形凸起相互配合可以腔体内部起密封作用，避免空气中的杂质进入到腔体内部，影响动物组织细胞生长情况。
优选的，所述的培养皿内壁上卡接有环形板，当培养皿内部动物组织细胞与培养液加入结束后将环形板卡接在培养皿内壁，避免培养皿滑动过程中培养液从培养皿中溅出，造成培养液的浪费，同时保证腔体内部的环境卫生。
优选的，相邻所述的调节凹槽的深度不同，使得竖直移动块在上下移动过程中获得不同振幅，进而通过晃动杆带动承托盘获得不同摇晃幅度，从而保证培养皿获得不同摇晃幅度使得培养液内部营养成分分布更加均匀。
优选的，所述的动物细胞组织培养装置对动物细胞的培养方法，包括以下步骤：第一步：放置处理：向装有动物组织细胞的培养皿内部加入培养液，将盖板取下后再将培养皿卡接在承托盘内部，最后将盖板合上。
第二步：晃动处理：晃动电机启动通过齿轮传动方式带动转轴转动，转轴通过转动圆盘带动竖直移动块在环形调节板上上下往复移动，进而竖直移动块通过晃动杆带动承托盘转动过程中同步带动承托盘晃动，从而对培养皿中的培养液进行晃动摇匀处理。
第三步：上料处理：当培养皿中的培养液不足时，打开密封塞经过进料口向上料筒内部灌入培养液，再将密封塞合上，根据各个培养皿中所需要培养液的多少，打开阀门通过注射筒挤压空气将培养液通过分液管与连接管相互配合将培养液加入培养皿中。
第四步：观察取出：通过显微放大镜观察动物组织细胞的生长情况，当动物组织细胞培养结束后打开盖板将培养皿取出，工作结束。
本发明的有益效果在于：1.本发明设计的培养框中分为四个腔体可以在动物细胞组织传代培养过程中同时对四个培养皿中的动物细胞组织进行增殖培养，提高动物细胞组织增殖培养效率；在晃动机构中，转动圆盘带动竖直移动块在环形调节板上上下往复移动，进而竖直移动块通过晃动杆带动承托盘转动过程中同步带动承托盘晃动，从而对培养皿中的培养液进行晃动摇匀处理；在上料机构中，打开密封塞经过进料口向上料筒内部灌入培养液，再将密封塞合上，根据各个培养皿中所需要培养液的多少，打开阀门通过注射筒挤压空气将培养液通过分液管与连接管相互配合将培养液泵入培养皿中，在整个上料过程中未打开盖板，从而减少杂菌进入腔体内部的可能，减少杂菌污染动物组织细胞培养的可能性。
2.本发明设计的上料机构中，当各个培养皿中需要的培养液相同时，同步打开阀门通过注射筒挤压空气将培养液通过分液管与连接管相互配合将培养液泵入培养皿中，当各个培养皿中所需要培养液不同时，分别打开阀门通过注射筒挤压空气将培养液通过分液管与连接管相互配合将培养液泵入培养皿中，通过调节阀门既可以同步向培养皿中泵入培养液，又可以分别向培养皿中泵入培养液，适用性更广。
3.本发明设计的上料筒中，当推动注射筒伸缩端下移时，此时滑动板同步下移，滑动板通过齿条带动传动齿轮转动，进而传动齿轮带动联动轴转动，联动轴通过一号锥齿轮与二号锥齿轮相互配合带动转动轴转动，从而转动轴带动搅拌爪对培养液搅匀处理，避免培养液在灌入上料筒内部后由于其长时间处于静置状态可能会产生营养成分沉淀的问题，提高培养液内部营养物质的均匀性。
 附图说明
下面结合附图和实施例对本发明进一步说明。
图1是本发明立体图。
图2是本发明主视图。
图3是本发明培养框内部立体结构示意图。
图4是本发明腔体与晃动机构内部安装结构示意图。
图5是本发明图4的A处局部剖视图。
图6是本发明腔体与晃动机构之间立体安装结构示意图。
图7是本发明上料筒内部结构示意图。
图8是本发明连接管内部结构示意图。
图9是本发明图8的B处局部放大图。
图10是本发明引流板立体结构示意图。
图11是本发明动物细胞组织培养流程图。
图中，1、培养平台；2、连接柱；3、培养框；31、密封槽；32、矩形凸起；4、盖板；41、显微放大镜；42、观察窗；5、晃动机构；51、晃动电机；52、转轴；53、转动圆盘；54、环形调节板；541、调节凹槽；55、竖直移动块；56、竖直连轴；561、贯通槽；57、晃动杆；58、转动套筒；59、移动连块；6、承托盘；61、环形件；62、承托弹簧杆；63、承托块；7、培养皿；71、环形板；8、上料机构；81、上料柱；82、上料筒；821、转动轴；822、搅拌爪；823、一号锥齿轮；824、联动轴；825、二号锥齿轮；826、传动齿轮；827、滑动板；828、复位弹簧杆；83、注射筒；84、分液管；85、连接管；851、导引板；852、引流板；853、引流槽；854、连接弹簧；855、擦拭海绵；856、导引绳；86、阀门；87、密封塞。
 具体实施方式
以下结合附图对本发明的实施例进行详细说明，但是本发明可以由权利要求限定和覆盖的多种不同方式实施。
如图1、图3以及图4所示，一种动物细胞组织培养装置，包括培养平台1、连接柱2、培养框3、盖板4、晃动机构5、承托盘6、培养皿7以及上料机构8，所述的培养平台1上端面四个拐角处安装有连接柱2，四个连接柱2上端面共同安装有培养框3，培养框3内部通过十字形隔板分隔成四个腔体，四个腔体内部以及培养平台1上端面共同安装有晃动机构5，晃动机构5上端安装有承托盘6，承托盘6上卡接有培养皿7，培养框3上端面安装有盖板4，盖板4上端面安装有上料机构8，盖板4上端面且位于腔体的正上方安装有观测腔体的显微放大镜41，培养框3侧壁四个拐角处开设有观察窗42。
如图3以及图4所示，所述的盖板4下端面边缘处设置有矩形凸起32，培养框3上端面边缘开设有与矩形凸起32相配合的密封槽31，密封槽31与矩形凸起32相互配合可以对腔体内部起密封作用，避免空气中的杂质进入到腔体内部，影响动物组织细胞生长情况。
如图6所示，所述的培养皿7内壁上卡接有环形板71，当培养皿7内部动物组织细胞与培养液加入结束后将环形板71卡接在培养皿7内壁，避免培养皿7晃动过程中培养液从培养皿7中溅出，造成培养液的浪费，同时保证腔体内部的环境卫生。
如图2、图4、图5以及图6所示，所述的晃动机构5包括晃动电机51、转轴52、转动圆盘53、环形调节板54、竖直移动块55、竖直连轴56、晃动杆57以及转动套筒58，其中所述的晃动电机51通过电机座安装在培养平台1上端面中部，培养框3下端且位于四个腔体中部通过轴承贯穿安装有转轴52，晃动电机51输出轴与四个转轴52之间均通过齿轮啮合方式相互连接，转轴52上端面安装有转动圆盘53，腔体底部中间位置安装有环形调节板54，环形调节板54上端面周向均匀开设有多个调节凹槽541，转动圆盘53上端上下贯穿滑动设置有竖直移动块55，且竖直移动块55下端抵靠在环形调节板54上，转动圆盘53上端面中部通过轴承安装有竖直连轴56，竖直连轴56上端面通过球铰安装有承托盘6，竖直连轴56中部自左向右开设有贯通槽561，贯通槽561内部通过铰轴安装有转动套筒58，转动套筒58内部滑动设置有晃动杆57，承托盘6下端面以及竖直移动块55上端面均水平滑动设置有移动连块59，晃动杆57上端通过铰链安装在上方的移动连块59上，晃动杆57下端通过铰链安装在下方的移动连块59上。
如图4以及图6所示，相邻所述的调节凹槽541的深度不同，使得竖直移动块55在上下移动过程中获得不同振幅，进而通过晃动杆57带动承托盘6获得不同摇晃幅度，从而保证培养皿7获得不同摇晃幅度使得培养液内部营养成分分布更加均匀。
如图1所示，所述的上料机构8包括上料柱81、上料筒82、注射筒83、分液管84以及连接管85，其中所述的上料柱81安装在盖板4上端面中部，上料筒82通过螺纹连接方式安装在上料柱81上端，上料筒82左侧开设有安装密封塞87的进料口，上料筒82上端面安装有与上料筒82相贯通且带有刻度的注射筒83，上料筒82下端面周向均匀安装有多个与上料筒82相贯通的分液管84，盖板4上端面且位于四个培养皿7正上方也安装有与腔体相贯通的分液管84，上下两个相邻分液管84之间卡接有带有阀门86的连接管85。
具体工作时，通过人工向装有动物组织细胞的培养皿7中加入培养液，将盖板4取下后再将培养皿7卡接在承托盘6内部，最后将盖板4合上，此时动物组织细胞在培养皿7内部增殖生长，通过显微放大镜41与观察窗42观察动物组织细胞的生长情况，由于动物组织细胞在增殖生长过程中需要消耗培养液中的营养成分，培养液中营养成分浓度自培养皿7中部向培养皿7四周逐渐减小，导致培养液内部营养成分不均，影响动物组织细胞生长，故在动物组织细胞培养过程中晃动电机51启动通过齿轮啮合方式带动转轴52转动，转轴52通过转动圆盘53带动竖直移动块55同步转动，竖直移动块55通过晃动杆57带动承托盘6绕着竖直连轴56转动，承托盘6转动过程中竖直移动块55通过与调节凹槽541相互配合带动晃动杆57在转动套筒58内部上下往复滑动，进而在承托盘6转动过程中通过晃动杆57同步带动承托盘6晃动，从而将培养皿7中的培养液晃动摇匀，避免培养液内部营养成分存在不均匀的问题，影响动物组织细胞增殖效果，其中上下两个移动连块59可以补偿晃动杆57在水平方向的位移。
当培养皿7中的培养液不足时，打开密封塞87经过进料口向上料筒82内部灌入培养液，再将密封塞87合上，根据各个培养皿7中所需要培养液的多少，当各个培养皿7中需要的培养液相同时，同步打开阀门86通过注射筒83向上料筒82挤压空气，挤压的空气通过分液管84与连接管85相互配合将培养液泵入培养皿7中，当各个培养皿7中所需要培养液不同时，分别打开阀门86通过注射筒83挤压空气将培养液通过分液管84与连接管85相互配合将培养液泵入培养皿7中，避免通过人工打开盖板4直接向培养皿7中加入培养液过程中引入杂菌的可能性，其中注射筒83上标有刻度可以实现向培养皿7中定量加入培养液，上料筒82与连接管85通过可拆卸方式分别安装在上料柱81与分液管84上，便于上料筒82与连接管85后续的快速拆卸清洗。
如图7所示，所述的上料筒82内部下端面中部通过轴承安装有转动轴821，转动轴821圆周面下方均匀设置有多个搅拌爪822，转动轴821上方安装有一号锥齿轮823，上料筒82右侧壁上方通过轴承安装有联动轴824，联动轴824左端面安装有与一号锥齿轮823相啮合的二号锥齿轮825，联动轴824上安装有传动齿轮826，上料筒82上端上下贯穿设置有滑动板827，滑动板827上端抵靠在注射筒83伸缩端下端面，滑动板827前端面下端设置有与传动齿轮826相配合的齿条，滑动板827底部设置有水平的固定凸起，滑动板827通过固定凸起与上料筒82内壁之间安装有复位弹簧杆828，培养液在灌入上料筒82内部后由于培养液处于静置状态，培养液内部的营养成分可能会发生沉淀的现象导致培养液内部存在营养物质不均匀的问题，故当按压注射筒83向培养皿7中泵入培养液时，此时滑动板827同步下移，滑动板827通过齿条带动传动齿轮826转动，进而传动齿轮826带动联动轴824转动，联动轴824通过一号锥齿轮823与二号锥齿轮825相互配合带动转动轴821转动，从而转动轴821带动搅拌爪822对培养液进行搅匀处理，提高培养液内部营养物质的均匀性，其中复位弹簧杆828对滑动板827起复位作用。
如图3以及图6所示，四个所述的腔体内壁且位于承托盘6外侧均安装有内部为环形件61，且环形件61为方形板上端面开设有圆通槽的结构，环形件61内壁上均匀安装有多个承托弹簧杆62，承托弹簧杆62远离环形件61的一端通过球铰安装有与承托盘6相配合的承托块63，环形件61通过承托弹簧杆62与承托块63相互配合可以对承托盘6起限位承托作用，避免承托盘6在竖直连轴56上单点受力不稳定性，增加承托盘6在晃动过程中稳定效果。
如图8、图9以及图10所示，所述的连接管85内部均匀设置多个开设有导引孔的导引板851，盖板4下端面且位于下方的分液管84出口处通过支撑架安装有引流板852，引流板852上端面设置为锥形结构，引流板852下端设置为伞型结构，伞型结构上端面周向均匀设置有多个引流槽853，下端为伞形结构的引流板852通过与引流槽853相互配合可以对培养液起到导流作用，避免培养液在进入培养皿7时分布不均匀，造成动物组织细胞获得营养成分不均匀的问题，影响动物组织细胞的增殖效果，导引板851可以减小培养液进入培养皿7中的冲击力，避免培养液的冲击力过大冲散动物组织细胞，影响动物组织细胞的培养情况。
如图9所示，所述的导引板851与连接管85内壁之间上下对称安装有连接弹簧854，导引板851与连接管85内壁接触的部分设置有擦拭海绵855，位于阀门86同一侧相邻两个导引板851之间安装有导引绳856，由于连接管85直径较小且较长，故在对连接管85通水进行清洗过程中手动拉动连接管85最外侧的两个导引板851移动，进而两个导引板851通过导引绳856带动剩余的导引板851在连接管85内部往复滑动，从而导引板851通过擦拭海绵855可以对连接管85内壁进行往复擦拭清洗，便于对连接管85内部的清理，其中连接弹簧854在清洗过程中对导引板851起复位作用。
如图11所示，本发明还提供了一种动物细胞的培养方法，包括以下步骤：第一步：放置处理：向装有动物组织细胞的培养皿7内部加入培养液，将盖板4取下后再将培养皿7卡接在承托盘6内部，最后将盖板4合上。
第二步：晃动处理：晃动电机51启动通过齿轮啮合方式带动转轴52转动，转轴52通过转动圆盘53带动竖直移动块55同步转动，竖直移动块55通过晃动杆57带动承托盘6绕着竖直连轴56转动，承托盘6转动过程中竖直移动块55通过与调节凹槽541相互配合带动晃动杆57在转动套筒58内部上下往复滑动，进而在承托盘6转动过程中通过晃动杆57同步带动承托盘6晃动，从而将培养皿7中的培养液晃动摇匀。
第三步：上料处理：当培养皿7中的培养液不足时，打开密封塞87经过进料口向上料筒82内部灌入培养液，再将密封塞87合上，根据各个培养皿7中所需要培养液的多少，打开阀门86通过注射筒83向上料筒82挤压空气，挤压的空气通过分液管84与连接管85相互配合将培养液泵入培养皿7中，注射筒83按压过程中此时滑动板827同步下移，滑动板827通过齿条带动传动齿轮826转动，进而传动齿轮826带动联动轴824转动，联动轴824通过一号锥齿轮823与二号锥齿轮825相互配合带动转动轴821转动，从而转动轴821带动搅拌爪822对培养液搅匀处理。
第四步：观察取出：通过显微放大镜41观察动物组织细胞的生长情况，当动物组织细胞培养结束后打开盖板4将培养皿7取出，工作结束。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (9)

  1. 一种动物细胞组织培养装置，包括培养平台(1)、连接柱(2)、培养框(3)、盖板(4)、晃动机构(5)、承托盘(6)、培养皿(7)以及上料机构(8)，其特征在于：所述的培养平台(1)上端面四个拐角处安装有连接柱(2)，四个连接柱(2)上端面共同安装有培养框(3)，培养框(3)内部通过十字形隔板分隔成四个腔体，四个腔体内部以及培养平台(1)上端面共同安装有晃动机构(5)，晃动机构(5)上端安装有承托盘(6)，承托盘(6)上卡接有培养皿(7)，培养框(3)上端面安装有盖板(4)，盖板(4)上端面安装有上料机构(8)，盖板(4)上端面且位于腔体的正上方安装有观测腔体的显微放大镜(41)，培养框(3)侧壁四个拐角处开设有观察窗(42)，其中：
    所述的晃动机构(5)包括晃动电机(51)、转轴(52)、转动圆盘(53)、环形调节板(54)、竖直移动块(55)、竖直连轴(56)、晃动杆(57)以及转动套筒(58)，其中所述的晃动电机(51)通过电机座安装在培养平台(1)上端面中部，培养框(3)下端且位于四个腔体中部通过轴承贯穿安装有转轴(52)，晃动电机(51)输出轴与四个转轴(52)之间均通过齿轮啮合方式相互连接，转轴(52)上端面安装有转动圆盘(53)，腔体底部中间位置安装有环形调节板(54)，环形调节板(54)上端面周向均匀开设有多个调节凹槽(541)，转动圆盘(53)上端上下贯穿滑动设置有竖直移动块(55)，且竖直移动块(55)下端抵靠在环形调节板(54)上，转动圆盘(53)上端面中部通过轴承安装有竖直连轴(56)，竖直连轴(56)上端面通过球铰安装有承托盘(6)，竖直连轴(56)中部自左向右开设有贯通槽(561)，贯通槽(561)内部通过铰轴安装有转动套筒(58)，转动套筒(58)内部滑动设置有晃动杆(57)，承托盘(6)下端面以及竖直移动块(55)上端面均水平滑动设置有移动连块(59)，晃动杆(57)上端通过铰链安装在上方的移动连块(59)上，晃动杆(57)下端通过铰链安装在下方的移动连块(59)上；
    所述的上料机构(8)包括上料柱(81)、上料筒(82)、注射筒(83)、分液管(84)以及连接管(85)，其中所述的上料柱(81)安装在盖板(4)上端面中部，上料筒(82)通过螺纹连接方式安装在上料柱(81)上端，上料筒(82)左侧开设有安装密封塞(87)的进料口，上料筒(82)上端面安装有与上料筒(82)相贯通且带有刻度的注射筒(83)，上料筒(82)下端面周向均匀安装有多个与上料筒(82)相贯通的分液管(84)，盖板(4)上端面且位于四个培养皿(7)正上方也安装有与腔体相贯通的分液管(84)，上下两个相邻分液管(84)之间卡接有带有阀门(86)的连接管(85)。
  2. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：所述的上料筒(82)内部下端面中部通过轴承安装有转动轴(821)，转动轴(821)圆周面下方均匀设置有多个搅拌爪(822)，转动轴(821)上方安装有一号锥齿轮(823)，上料筒(82)右侧壁上方通过轴承安装有联动轴(824)，联动轴(824)左端面安装有与一号锥齿轮(823)相啮合的二号锥齿轮(825)，联动轴(824)上安装有传动齿轮(826)，上料筒(82)上端上下贯穿设置有滑动板(827)，滑动板(827)上端抵靠在注射筒(83)伸缩端下端面，滑动板(827)前端面下端设置有与传动齿轮(826)相配合的齿条，滑动板(827)底部设置有水平的固定凸起，滑动板(827)通过固定凸起与上料筒(82)内壁之间安装有复位弹簧杆(828)。
  3. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：四个所述的腔体内壁且位于承托盘(6)外侧均安装有环形件(61)，且环形件(61)为方形板上端面开设有圆通槽的结构，环形件(61)内壁上均匀安装有多个承托弹簧杆(62)，承托弹簧杆(62)远离环形件(61)的一端通过球铰安装有与承托盘(6)相配合的承托块(63)。
  4. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：所述的连接管(85)内部均匀设置多个开设有导引孔的导引板(851)，盖板(4)下端面且位于下方的分液管(84)出口处通过支撑架安装有引流板(852)，引流板(852)上端面设置为锥形结构，引流板(852)下端设置为伞型结构，伞型结构上端面周向均匀设置有多个引流槽(853)。
  5. 根据权利要求4所述的一种动物细胞组织培养装置，其特征在于：所述的导引板(851)与连接管(85)内壁之间上下对称安装有连接弹簧(854)，导引板(851)与连接管(85)内壁接触的部分设置有擦拭海绵(855)，位于阀门(86)同一侧相邻两个导引板(851)之间安装有导引绳(856)。
  6. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：所述的盖板(4)下端面边缘处设置有矩形凸起(32)，培养框(3)上端面边缘开设有与矩形凸起(32)相配合的密封槽(31)。
  7. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：所述的培养皿(7)内壁上卡接有环形板(71)。
  8. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：相邻所述的调节凹槽(541)的深度不同。
  9. 根据权利要求1所述的一种动物细胞组织培养装置，其特征在于：所述的动物细胞组织培养装置对动物细胞的培养方法，包括以下步骤：
    第一步：放置处理：向装有动物组织细胞的培养皿(7)内部加入营养液，将盖板(4)取下后再将培养皿(7)卡接在承托盘(6)内部，最后将盖板(4)合上；
    第二步：晃动处理：晃动电机(51)启动通过齿轮传动方式带动转轴(52)转动，转轴(52)通过转动圆盘(53)带动竖直移动块(55)在环形调节板(54)上上下往复移动，进而竖直移动块(55)通过晃动杆(57)带动承托盘(6)转动过程中同步带动承托盘(6)晃动，从而对培养皿(7)中的营养液进行晃动摇匀处理；
    第三步：上料处理：当培养皿(7)中的营养液不足时，打开密封塞(87)经过进料口向上料筒(82)内部灌入营养液，再将密封塞(87)合上，根据各个培养皿(7)中所需要营养液的多少，打开阀门(86)通过注射筒(83)挤压空气将营养液通过分液管(84)与连接管(85)相互配合将营养液加入培养皿(7)中；
    第四步：观察取出：通过显微放大镜(41)观察动物组织细胞的生长情况，当动物组织细胞培养结束后打开盖板(4)将培养皿(7)取出，工作结束。
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