CN112608847A - Sealed cultivation device for biological cell cultivation and use method thereof - Google Patents

Sealed cultivation device for biological cell cultivation and use method thereof Download PDF

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Publication number
CN112608847A
CN112608847A CN202110109524.4A CN202110109524A CN112608847A CN 112608847 A CN112608847 A CN 112608847A CN 202110109524 A CN202110109524 A CN 202110109524A CN 112608847 A CN112608847 A CN 112608847A
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culture dish
transparent
clamping plate
culture
sealed
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张海甄
郭莹莹
郭穗穗
易麗夏
梁欣儿
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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
    • C12M23/02—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
    • 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
    • C12M23/48—Holding appliances; Racks; Supports
    • 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
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • 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
    • C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
    • 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
    • C12M29/20—Degassing; Venting; Bubble traps
    • 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
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C12M37/04—Seals
    • 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
    • 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
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements

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  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
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  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention discloses a sealed cultivation device for cultivating biological cells, which comprises a clamping device, a transparent culture dish lower cover and a transparent culture dish upper cover, wherein the transparent culture dish upper cover is covered above the transparent culture dish lower cover; the clamping device comprises a fork-shaped upper clamping plate and a fork-shaped lower clamping plate, the upper clamping plate and the lower clamping plate are in threaded connection with threaded rods and are in sliding connection with sliding guide rods; the threaded rod comprises a positive thread part in threaded connection with the upper clamping plate and a reverse thread part in threaded connection with the lower clamping plate. The invention realizes the automatic inspection of the culture medium, the automatic culture medium feeding, the oxygen introduction and the vibration during the cell culture, effectively reduces the workload of scientific research personnel and improves the working efficiency.

Description

Sealed cultivation device for biological cell cultivation and use method thereof
Technical Field
The invention belongs to the field of biology, and particularly relates to a sealed cultivation device for cultivating biological cells and a using method thereof.
Background
When organisms need to be cultured, sealed culture is usually required to avoid external bacterial infection. However, in the case of sealed culture, it is usually necessary to supplement the culture medium depending on the degree of growth of the cells. This requires frequent opening and closing of the culture apparatus, which is likely to cause contamination of the culture medium. In addition, the culture device needs to be turned over manually and periodically to prevent the cells from stopping growing after being attached to the wall, so that improvement is needed.
Disclosure of Invention
In order to solve the above problems, the present invention provides a sealed cultivation device for cultivating biological cells and a method of using the same. The invention realizes the automatic inspection of the culture medium, the automatic culture medium feeding, the oxygen introduction and the vibration during the cell culture, effectively reduces the workload of scientific research personnel and improves the working efficiency.
In order to achieve the technical effects, the technical scheme of the invention is as follows:
a sealed cultivation device for cultivating biological cells comprises a clamping device, a transparent culture dish lower cover and a transparent culture dish upper cover covering the transparent culture dish lower cover; the clamping device comprises a fork-shaped upper clamping plate and a fork-shaped lower clamping plate, the upper clamping plate and the lower clamping plate are in threaded connection with threaded rods and are in sliding connection with sliding guide rods; the threaded rod comprises a positive thread part in threaded connection with the upper clamping plate and a reverse thread part in threaded connection with the lower clamping plate; the threaded rod is connected with a first rotating device, and the bottom of the sliding guide rod is connected with the first rotating device through a connecting block; the middle part of the sliding guide rod is connected with a vibration motor; the upper cover of the transparent culture dish is communicated with a culture solution storage box through a first hose; the first hose is sequentially provided with a first flowmeter, a liquid feeding pump and a first one-way valve from top to bottom; the upper cover of the transparent culture dish is communicated with an oxygen generator through a second hose, and a second flowmeter and a second one-way valve are sequentially arranged on the second hose from top to bottom; the upper cover of the transparent culture dish is also communicated with an air outlet pipe, and a waterproof breathable film and a third one-way valve are arranged on the air outlet pipe; a machine vision system is arranged above the upper cover of the transparent culture dish or below the lower cover of the transparent culture dish; the machine vision system is wirelessly connected with the liquid feeding pump; the vibration motor and the oxygen generator are respectively and electrically connected with a timer.
In a further improvement, the top of the air outlet pipe is also plugged with a sterile surface.
In a further improvement, an annular groove matched with the lower cover of the transparent culture dish is formed on the upper cover of the transparent culture dish, and an elastic sealing gasket is fixed on the annular groove.
In a further improvement, the top and the bottom of the sliding guide rod are respectively fixed with a baffle plate.
In a further improvement, the first rotating device is a rotating electric machine.
In a further improvement, positioning grooves are formed on the lower cover of the transparent culture dish and the upper cover of the culture dish, and corresponding positioning bosses are formed on the upper clamping plate and the lower clamping plate.
A method for using a sealed cultivation device for cultivating biological cells, comprising the following steps:
step one, installing the sealed culture device in a sterile incubator, and then injecting cultured cells and a culture medium into a lower cover of a culture dish; covering the transparent culture dish;
step two, the machine vision system photographs the culture dish to obtain an initial picture; then, the machine vision system photographs at preset time intervals and compares the photographs to obtain the cell concentration in the culture dish; the vibration motor vibrates at each preset time, and the oxygen generator charges oxygen at each preset time;
and step three, when the machine vision system senses that the cell concentration reaches the concentration of the culture solution to be added, controlling the liquid feeding pump to start, and injecting the culture solution into the culture dish.
And step four, after the total volume of the injected culture solution reaches a preset volume and the cell concentration reaches a preset concentration, the machine vision system sends information to a mobile phone of a culturist to remind the culturist.
Taking the lower cover of the transparent culture dish down to extract the cells; then, each apparatus was sterilized and cultured next time.
The invention has the advantages that:
the invention realizes the automatic inspection of the culture medium, the automatic culture medium feeding, the oxygen introduction and the vibration during the cell culture, effectively reduces the workload of scientific research personnel and improves the working efficiency.
Drawings
Fig. 1 is a schematic view of the overall structure of the present invention.
Detailed Description
The technical means of the present invention will be specifically described below by way of specific embodiments.
Example 1
A sealed culture device for culturing biological cells as shown in FIG. 1 comprises a holding device 1, a transparent culture dish lower cover 2 and a transparent culture dish upper cover 3 covering the transparent culture dish lower cover 2; the clamping device 1 comprises a fork-shaped upper clamping plate 4 and a fork-shaped lower clamping plate 5, wherein the upper clamping plate 4 and the lower clamping plate 5 are in threaded connection with a threaded rod 6 and are in sliding connection with a sliding guide rod 7; the threaded rod 6 comprises a positive thread part 8 in threaded connection with the upper clamping plate 4 and a reverse thread part 9 in threaded connection with the lower clamping plate 5; the threaded rod 6 is connected with a first rotating device 10, and the bottom of the sliding guide rod 7 is connected with the first rotating device 10 through a connecting block 11; the middle part of the sliding guide rod 7 is connected with a vibration motor 12; the transparent culture dish upper cover 3 is communicated with a culture solution storage box 31 through a first hose 13; the first hose 13 is provided with a first flowmeter 14, a liquid feeding pump 15 and a first one-way valve 16 from top to bottom in sequence; the upper cover 3 of the transparent culture dish is communicated with an oxygen generator 18 through a second hose 17, and a second flowmeter 19 and a second one-way valve 20 are sequentially arranged on the second hose 17 from top to bottom; the upper cover 3 of the transparent culture dish is also communicated with an air outlet pipe 21, and a waterproof breathable film 22 and a third one-way valve 23 are arranged on the air outlet pipe 21; a machine vision system 28 is arranged above the upper cover 3 of the transparent culture dish or below the lower cover 2 of the transparent culture dish; the machine vision system 28 is wirelessly connected with the liquid sending pump 15; the oscillation motor 12 and the oxygen generator 18 are electrically connected with a timer respectively. The top of the air outlet pipe 21 is also plugged with a sterile surface 24.
An annular groove 25 matched with the transparent culture dish lower cover 2 is formed on the transparent culture dish upper cover 3, and an elastic sealing gasket 26 is fixed on the annular groove 25. The slide guide 7 is fixed at the top and bottom thereof with stoppers 27.
The first rotating device 10 is a rotating electrical machine.
The transparent culture dish lower cover 2 and the culture dish upper cover 3 are both provided with positioning grooves 29, and the upper clamping plate 4 and the lower clamping plate 5 are provided with corresponding positioning convex blocks 30.
The use method of the sealed culture device for culturing the biological cells comprises the following steps:
step one, installing the sealed culture device in a sterile incubator, and then injecting cultured cells and a culture medium into a lower cover 2 of a culture dish; covering a transparent culture dish upper cover 3;
step two, the machine vision system 28 takes a picture of the culture dish to obtain an initial picture; then the machine vision system 28 takes pictures at preset time intervals and compares the pictures to obtain the cell concentration in the culture dish; the oscillation motor 12 vibrates at each preset time, and the oxygen generator 18 is charged with oxygen at each preset time;
and step three, when the machine vision system 28 senses that the cell concentration reaches the concentration of the culture solution required to be added, controlling the liquid feeding pump 15 to start, and injecting the culture solution into the culture dish.
Step four, after the total volume of the injected culture solution reaches the preset volume and the cell concentration reaches the preset concentration, the machine vision system 28 sends information to the mobile phone of the culturist to remind the culturist.
Step five, taking down the lower cover 2 of the transparent culture dish to extract cells in the culture dish by a culture person; then, each apparatus was sterilized and cultured next time.
The above description is only one specific guiding embodiment of the present invention, but the design concept of the present invention is not limited thereto, and any insubstantial modification of the present invention using this concept shall fall within the scope of the invention.

Claims (7)

1. A sealed cultivation device for biological cell cultivation is characterized by comprising a clamping device (1), a transparent culture dish lower cover (2) and a transparent culture dish upper cover (3) covering the transparent culture dish lower cover (2); the clamping device (1) comprises a fork-shaped upper clamping plate (4) and a fork-shaped lower clamping plate (5), the upper clamping plate (4) and the lower clamping plate (5) are in threaded connection with a threaded rod (6) and are in sliding connection with a sliding guide rod (7); the threaded rod (6) comprises a positive thread part (8) in threaded connection with the upper clamping plate (4) and a reverse thread part (9) in threaded connection with the lower clamping plate (5); the threaded rod (6) is connected with a first rotating device (10), and the bottom of the sliding guide rod (7) is connected with the first rotating device (10) through a connecting block (11); the middle part of the sliding guide rod (7) is connected with a vibration motor (12); the upper cover (3) of the transparent culture dish is communicated with a culture solution storage box (31) through a first hose (13); the first hose (13) is sequentially provided with a first flowmeter (14), a liquid feeding pump (15) and a first one-way valve (16) from top to bottom; the upper cover (3) of the transparent culture dish is communicated with an aerobic generator (18) through a second hose (17), and a second flowmeter (19) and a second one-way valve (20) are sequentially arranged on the second hose (17) from top to bottom; the upper cover (3) of the transparent culture dish is also communicated with an air outlet pipe (21), and a waterproof breathable film (22) and a third one-way valve (23) are arranged on the air outlet pipe (21); a machine vision system (28) is arranged above the upper cover (3) of the transparent culture dish or below the lower cover (2) of the transparent culture dish; the machine vision system (28) is wirelessly connected with the liquid sending pump (15); the oscillation motor (12) and the oxygen generator (18) are respectively and electrically connected with a timer.
2. A sealed culturing device for culturing biological cells, according to claim 1, wherein the top of said air outlet tube (21) is plugged with a sterile surface (24).
3. The sealed culture device for culturing biological cells according to claim 2, wherein the transparent upper lid (3) of the culture dish is formed with an annular groove (25) that is engaged with the lower lid (2) of the culture dish, and an elastic packing (26) is fixed to the annular groove (25).
4. The sealed culturing device for culturing biological cells according to claim 1, wherein the sliding guide bar (7) has a stopper (27) fixed to both the top and bottom thereof.
5. The sealed culturing device for culturing biological cells according to claim 1, wherein the first rotating means (10) is a rotating motor.
6. The sealed culturing device for culturing biological cells according to claim 1, wherein the transparent lower cover (2) and the upper cover (3) of the culture dish are each formed with a positioning groove (29), and the upper clamping plate (4) and the lower clamping plate (5) are each formed with a corresponding positioning protrusion (30).
7. A method for using a sealed culture device for culturing biological cells, comprising the steps of:
step one, the sealed culture device is arranged in a sterile incubator, and then culture cells and culture media are injected into a lower cover (2) of a culture dish; covering a transparent culture dish upper cover (3);
step two, the machine vision system (28) photographs the culture dish to obtain an initial picture; then a machine vision system (28) shoots at preset time intervals to detect the transparency of the culture medium, so as to correspondingly obtain the cell concentration in the culture dish; the oscillation motor (12) vibrates at each preset time, and the oxygen generator (18) is charged with oxygen at each preset time;
and step three, when the machine vision system (28) senses that the cell concentration reaches the concentration of the culture solution required to be added, controlling the liquid feeding pump (15) to start, and injecting the culture solution into the culture dish.
And step four, after the total volume of the injected culture solution reaches a preset volume and the cell concentration reaches a preset concentration, the machine vision system (28) sends information to a mobile phone of a culturist to remind the culturist.
Step five, taking down the lower cover (2) of the transparent culture dish by the culturist to extract the cells in the transparent culture dish; then, each apparatus was sterilized and cultured next time.
CN202110109524.4A 2021-01-27 2021-01-27 Sealed cultivation device for biological cell cultivation and use method thereof Pending CN112608847A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113528340A (en) * 2021-08-04 2021-10-22 河南省肿瘤医院 Tumor cell culture system
CN115125101A (en) * 2022-07-29 2022-09-30 广州堃盛医疗用品有限公司 Microbial fermentation detection device

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CN110106065A (en) * 2015-06-04 2019-08-09 泷源制造株式会社 Cell, tissue are transported with without swing shell
CN209493569U (en) * 2018-09-30 2019-10-15 山东和田旺生物科技有限公司 A kind of microorganism fungus kind concussion and cultivate case
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JP2005080606A (en) * 2003-09-10 2005-03-31 Bio Map Co Cell culture apparatus
US20130011224A1 (en) * 2010-03-18 2013-01-10 Quantifoil Instruments Gmbh Positioning unit for a functional unit
CN104169413A (en) * 2012-02-29 2014-11-26 株式会社Jtec Cell culture device having culture medium replacement function
CN110106065A (en) * 2015-06-04 2019-08-09 泷源制造株式会社 Cell, tissue are transported with without swing shell
US20190017009A1 (en) * 2015-12-23 2019-01-17 Shanghai GenBase Biotechnology Co., Ltd. Fully automated continuous cell culture system
CN105749791A (en) * 2016-04-19 2016-07-13 成都中牧生物药业有限公司 Uniform mixing device on basis of different fixed sizes
CN106047691A (en) * 2016-05-28 2016-10-26 上海大学 3D cell culture system
CN207159236U (en) * 2017-07-25 2018-03-30 内蒙古医科大学附属医院 A kind of cell culture apparatus for strengthening oxygen supply
CN108441411A (en) * 2018-04-02 2018-08-24 郭庆 A kind of aerobic Bacteria Culture system
CN108485968A (en) * 2018-04-28 2018-09-04 李晓伟 A kind of novel cytotostatic culture device convenient for observation
CN208545429U (en) * 2018-05-23 2019-02-26 江苏益玛生物科技有限公司 A kind of portable high-efficiency biological respinse culture dish
CN209493569U (en) * 2018-09-30 2019-10-15 山东和田旺生物科技有限公司 A kind of microorganism fungus kind concussion and cultivate case
CN109162993A (en) * 2018-11-05 2019-01-08 赵本博 A kind of clamping device
CN210419992U (en) * 2019-08-16 2020-04-28 赵雨轩 Cell culture device

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113528340A (en) * 2021-08-04 2021-10-22 河南省肿瘤医院 Tumor cell culture system
CN115125101A (en) * 2022-07-29 2022-09-30 广州堃盛医疗用品有限公司 Microbial fermentation detection device

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