CN113913295B - Immune cell CAR-T culture equipment with high cell survival rate and culture method - Google Patents
Immune cell CAR-T culture equipment with high cell survival rate and culture method Download PDFInfo
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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
- C12M23/06—Tubular
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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
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- 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
- C12M23/52—Mobile; Means for transporting the apparatus
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- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/12—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by pressure
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- 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
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- 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
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
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- 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
- C12M41/48—Automatic or computerized control
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- 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
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0636—T lymphocytes
Abstract
The invention relates to the technical field of cell culture, and provides immune cell CAR-T culture equipment with high cell survival rate and a culture method, the immune cell CAR-T culture equipment comprises an equipment outer frame, a culture rack and an adjusting rotary valve, wherein the equipment outer frame comprises an observation window, the culture rack comprises a culture inner chamber, the culture rack is arranged on the inner side of the equipment outer frame, a culture bin for culturing cells is arranged on the inner side of the culture inner chamber, the observation window is arranged at the top end of a reaction tube, the inner side of the culture bin is movably connected with the reaction tube for sensing the concentration of carbon dioxide in the interior, one side of the culture bin, which is far away from the reaction tube, is movably connected with a transmission tube for receiving nutrients, an auxiliary block drives an auxiliary clamping block to move outwards through a rotary rod column, and the auxiliary clamping block drives a sliding block to move outwards, so that staff can rapidly check the specific conditions of the cells in the interior through the observation window, thereby realizing the stock rate of the cells observed at any time and avoiding cell death.
Description
Technical Field
The invention relates to the technical field of cell culture, in particular to immune cell CAR-T culture equipment with high cell survival rate and a culture method.
Background
Chimeric antigen receptors (Chimeric Antigen Receptor, CARs) are artificial modified fusion proteins based on T cell receptors, consisting of an extracellular antigen recognition domain and an intracellular signaling domain. Chimeric antigen receptor T cells (CAR-T cells) are obtained by coupling the antigen binding portion of an antibody recognizing a tumor antigen to the CD3- ζ chain or the intracellular portion of fceriγ in vitro to form a chimeric protein, and transfecting T cells of a patient by gene transduction to express the Chimeric Antigen Receptor (CAR). Chimeric Antigen Receptor (CAR) consists of 4 parts of the extracellular antigen binding region (scFv), hinge region, intermediate transmembrane region and intracellular signaling region of T cell receptor. Currently, a variety of tumor antigens are available for extracellular domain recognition, including CD19, CD20, epidermal Growth Factor Receptor (EGFR), human epidermal growth factor receptor 2 (HER-2), and the like. The Hinge region includes CD8Hinge, CD28Hinge, and longer IgG1Fc or IgG4Fc, etc. The middle transmembrane region has CD4, CD8, CD28, etc., and anchors the CAR structure to the T cell membrane. The intracellular domain typically comprises cd3ζ, CD28,4-1BB or OX40 for increasing T cell activation. T cells genetically modified to express a CAR can directly recognize the antigen targeted by the CAR and then trigger tumors that express CAR-specific antigens.
Currently, CAR modified T cells have met with significant success in the treatment of hematological cancers, including lymphomas, chronic lymphocytic leukemia and Acute Lymphocytic Leukemia (ALL). CD 19-targeted CAR-T cells are reported to have a complete remission rate of 70% to 90% in all patients. Therefore, the CART cells have wide application prospect in tumor immunotherapy and huge market value.
However, the current immune cell CAR-T always needs a worker to observe the cultivation condition of the immune cell CAR-T at any time in the cultivation process, and for the transportation and specific conditions of the nutrient substances needed by the immune cell CAR-T, the specific cultivation condition of the immune cell CAR-T always needs to be observed for many times, the whole cultivation condition is slower, the immune cell CAR-T has unexpected conditions in the cultivation process and cannot be timely noticed, and the cell survival rate is lower, so that the immune cell CAR-T cultivation equipment and the immune cell CAR-T cultivation method with high cell survival rate are provided.
Disclosure of Invention
In order to solve the technical problems, the invention provides immune cell CAR-T culture equipment with high cell survival rate and a culture method, which are achieved by the following specific technical means:
the utility model provides an immune cell CAR-T cultivates equipment that cell survival rate is high, includes the equipment frame, cultivates the frame, the adjustment rotary valve, the equipment frame includes the observation window, cultivate the frame including cultivateing the inner room, cultivate the frame setting in the inboard of equipment frame, the inboard of cultivateing the inner room is provided with the cultivation storehouse that is used for cultivating the cell, the observation window sets up the top at the reaction tube, the inboard swing joint who cultivates the storehouse has the reaction tube that is used for feeling inside carbon dioxide concentration, the one side swing joint that cultivates the storehouse and keep away from the reaction tube has the transfer pipe that is used for receiving the nutrient, the outside swing joint of reaction tube has the gasbag piece that is used for conveying inside atmospheric pressure, the outside swing joint of reaction tube has central runner, the outside swing joint of gasbag piece has the deflection pole, the both sides of deflection pole are provided with curved cardboard, the adjustment rotary valve includes the transfer pipe, the outside swing joint of transfer pipe has the slip gasbag, the adjustment rotary valve sets up the bottom at cultivateing the frame, through the concrete condition of the inside cell of storehouse, carries out the analysis according to the reaction tube, will handle mode and divide into urgent processing nutrient interpolation and cultivation situation into normal situation.
Preferably, the both sides swing joint of equipment frame has the roll adjustment frame of being convenient for observe the cell, the outside of roll adjustment frame is provided with the sealing washer, the sealing washer of roll adjustment frame is the closed state setting with the equipment frame, and whole framework is comparatively simple, and prevents that outside bacterium from getting into, guarantees inside for aseptic condition.
Preferably, the distance-adjusting frame comprises an auxiliary clamping block, a rotating rod column used for deflecting is movably connected to the outer side of the auxiliary clamping block, an auxiliary block is movably connected to the outer side of the rotating rod column, and the sliding block is driven to be pulled out through the deflection of the rotating rod column, so that staff in a research institute can observe the distance-adjusting frame rapidly.
Preferably, the both sides swing joint of supplementary piece has the gear rack that is used for being convenient for to pull, the outside of gear rack is provided with and is used for spacing gasbag post, the both sides of gasbag post are provided with the spacing that is used for preventing to remove, prevent to be in the cultivation in-process, because remove and cause the cell to appear colliding the damage.
Preferably, the quantity of cultivating the storehouse is the multiunit, and the multiunit cultivate the storehouse and be ring array setting about central runner, the top of cultivating the storehouse is provided with the sealing block, and cultivates the inside aseptic environment that is of storehouse, guarantees inside suitable cultivation condition.
Preferably, the adjusting rotary valve is internally provided with a staged latch, the staged latch of the adjusting rotary valve and the sliding air bag are arranged in an extrudable state, the staged latch of the adjusting rotary valve is arranged in a deflection state under the pushing of the deflection rod, the deflection distance is limited, the working states of the adjusting rotary valve are separated, and the adjusting rotary valve is suitable for cultivation of cells.
Preferably, the lug has been seted up to the both sides of gasbag post, the lug and the spacing looks adaptation of gasbag post, the bottom of gear rack is provided with the rubber sleeve, the rubber sleeve of gear rack is sealed state setting with the gasbag post, the bottom swing joint of gear rack has the support frame, the top swing joint of support frame has the commentaries on classics tooth frame, guarantees the life state of inside cell, is convenient for observe.
Preferably, the number of the air bag blocks and the deflection rods is multiple, and the air bag blocks and the deflection rods are arranged in a circular ring shape relative to the central rotating wheel, so that multiple groups of cell culture and comparison observation can be realized.
An immune cell CAR-T culture method with high cell survival rate comprises the following steps:
s1: preparation process of self-adding nutrient
When the cells are cultivated in the cultivation bin, when more oxygen and carbon dioxide are consumed, the carbon dioxide is used for adjusting the internal PH, constant PH is regulated according to the requirements of the cultivated cells, most of PH is cultivated under the condition of PH7.2-7.4, the change of PH reacts with the reagent in the reaction tube to form an electric signal which is transmitted to the electric signal receiver, then the electric signal receiver is used for increasing the internal air pressure of the air bag block through instructions to form an air pressure difference, and the air pressure difference drives the deflection rod to rotate.
S2: nutrient addition step
And secondly, the deflection rod deflects the outer side of the rotary valve by extruding and adjusting the outer side of the rotary valve to a fixed distance, the deflection angle is 0-30 degrees, the rotary valve is adjusted to open a switch of internal air pressure, the rotary valve is adjusted to drive the sliding air bag to be extruded through sliding friction force between the rotary valve and the sliding air bag, and then the air pressure drives the internal nutrient to be transmitted to the inside of the cultivation bin through the transmission pipe.
S3: alarm step of cultivation
If the cells are in partial death or abnormal conditions in the cultivation process, the PH condition in the cultivation bin is greatly changed and is higher than PH7.6, so that the reaction with the reagent in the reaction tube is stronger, an electric signal is formed and transmitted to the electric signal receiver, the electric signal receiver drives the deflection rod to deflect to 30-60 degrees through the instruction, the curved clamping plate limits the moving distance of the electric signal receiver, and the deflection rod drives the adjusting rotary valve to rotate to an alarm area, so that the equipment gives an alarm.
S4: emergency observation step
Excessive deflection of the adjusting rotary valve drives the gear rack to move backwards, the gear rack and the supporting frame limit the deflection angle of the gear rack, the gear rack drives the auxiliary block to move outwards, the auxiliary block drives the auxiliary clamping block to move outwards through the rotating rod column, the auxiliary clamping block drives the sliding block to move outwards, and then workers can rapidly check the specific conditions of internal cells through the observation window.
The invention has the following beneficial effects:
1. according to the immune cell CAR-T culture equipment with high cell survival rate and the culture method, when cells are cultured in a culture bin, more oxygen and carbon dioxide are consumed, the carbon dioxide is used for adjusting the internal PH, constant PH is regulated according to the requirement of the cultured cells, most PH is cultured under the condition of PH7.2-7.4, the change of PH reacts with the reagent in a reaction tube to form an electric signal which is transmitted to an electric signal receiver, the electric signal receiver increases the internal air pressure of an air bag block through instructions to form air pressure difference, the air pressure difference drives a deflection rod to rotate, the deflection rod is used for adjusting the outer side of a rotary valve through extrusion to deflect a fixed distance, the deflection angle is 0-30 degrees, the rotary valve is adjusted to open a switch of the internal air pressure, and the rotary valve is adjusted to drive the sliding air bag to be extruded through sliding friction force between the rotary valve and the sliding air bag, so that the air pressure drives the internal nutrient to be transmitted to the culture bin through a transmission tube, and the automatic nutrient conveying is realized, and a worker does not need to pay attention to specific culture conditions at any time, and the whole culture condition is relatively large.
2. According to the immune cell CAR-T culture equipment and the culture method with high cell survival rate, the air bag block drives the deflection rod to deflect to the degree, the curved clamping plate limits the moving distance of the immune cell CAR-T culture equipment, the deflection rod drives the adjustment rotary valve to rotate to the alarm area, the equipment gives an alarm, excessive deflection of the adjustment rotary valve drives the gear rack to move backwards, the gear rack and the support frame limit the deflection angle of the gear rack, the gear rack drives the auxiliary block to move outwards, the auxiliary block drives the auxiliary clamping block to move outwards through the rotating rod column, the auxiliary clamping block drives the sliding block to move outwards, and then workers can rapidly check specific conditions of internal cells through the observation window, so that the stock rate of the cells is observed at any time, and accidental death of the cells is avoided.
Drawings
FIG. 1 is a schematic diagram of the outer frame structure of the device of the present invention;
FIG. 2 is a schematic view of a turning bar structure according to the present invention;
FIG. 3 is a partial cross-sectional view of FIG. 2 of the present invention;
FIG. 4 is a schematic view of the structure of a reaction tube according to the present invention;
FIG. 5 is a partial cross-sectional view of FIG. 4 in accordance with the present invention;
FIG. 6 is a schematic view of a cultivation frame structure according to the present invention;
fig. 7 is a schematic structural view of a support frame according to the present invention.
In the figure: 1. an equipment outer frame; 101. an observation window; 2. a distance-adjusting frame; 201. an auxiliary block; 202. a rotating rod column; 203. an auxiliary clamping block; 204. a sliding block; 205. a gear rack; 206. an airbag column; 207. a limiting frame; 208. a support frame; 209. a rotating rack; 3. a cultivation rack; 301. cultivating an inner room; 302. a cultivation bin; 303. a reaction tube; 304. a curved clamping plate; 305. a deflection lever; 306. an airbag block; 307. a central rotating wheel; 4. adjusting a rotary valve; 401. a transfer tube; 402. and a sliding air bag.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to fig. 1-5, embodiment one: the utility model provides an immune cell CAR-T cultivates equipment that cell survival rate is high, including equipment frame 1, cultivate frame 3, adjust rotary valve 4, equipment frame 1 includes observation window 101, cultivate frame 3 and including cultivateing inner room 301, cultivate the inboard of frame 3 setting at equipment frame 1, cultivate the inboard of inner room 301 and be provided with the cultivation storehouse 302 that is used for cultivating the cell, the observation window 101 sets up the top at reaction tube 303, the inboard swing joint of cultivation storehouse 302 has the reaction tube 303 that is used for feeling inside carbon dioxide concentration, one side swing joint that reaction tube 303 was kept away from to cultivation storehouse 302 has the transfer pipe 401 that is used for receiving the nutrient, the outside swing joint of reaction tube 303 has the gasbag piece 306 that is used for transmitting inside atmospheric pressure, the outside swing joint of reaction tube 303 has central runner 307, the outside swing joint of gasbag piece 306 has deflector 305, the both sides of deflector 305 are provided with curved cardboard 304.
Referring to fig. 3-5, the adjusting rotary valve 4 includes a transfer tube 401, a sliding air bag 402 is movably connected to the outer side of the transfer tube 401, the adjusting rotary valve 4 is disposed at the bottom end of the cultivating frame 3, by analyzing the specific condition of cells in the cultivating bin 302 according to the reaction tube 303, the processing mode is divided into normal nutrient adding cultivating and emergency processing conditions, the number of the cultivating bins 302 is multiple groups, the cultivating bins 302 are disposed in a circular array with respect to the central rotating wheel 307, a sealing block is disposed at the top end of the cultivating bins 302, the inside of the cultivating bins 302 is in a sterile environment, suitable cultivating conditions in the interior are ensured, the number of the air bag blocks 306 and the deflecting rods 305 is multiple groups, the air bag blocks 306 and the deflecting rods 305 are disposed in a circular ring shape with respect to the central rotating wheel 307, multiple groups of cell cultivation and comparison observation can be realized, the staged latch teeth of the adjusting rotary valve 4 are disposed in a squeezable state with the sliding air bag 402, the staged latch of the adjusting rotary valve 4 is disposed in a deflecting state under the pushing of the deflecting rods 305, and the deflecting distance is limited, and the working state of the adjusting rotary valve 4 is adapted to the cultivating cells.
Referring to fig. 6-7, embodiment two: the both sides swing joint of equipment frame 1 has the roll adjustment frame 2 of being convenient for observe the cell, the outside of roll adjustment frame 2 is provided with the sealing washer, the sealing washer of roll adjustment frame 2 is the closed state setting with equipment frame 1, the overall framework is comparatively simple, and prevent outside bacterium entering, guarantee inside aseptic condition, roll adjustment frame 2 includes auxiliary fixture block 203, the outside swing joint of auxiliary fixture block 203 has the bull stick post 202 that is used for deflecting, the outside swing joint of bull stick post 202 has auxiliary block 201, pull out through the bull stick post 202 drives slider 204, cause the institute personnel to observe fast, the both sides swing joint of auxiliary block 201 has the rack 205 that is used for being convenient for pull, the outside of rack 205 is provided with and is used for spacing gasbag post 206, the both sides of gasbag post 206 are provided with the spacing frame 207 that is used for preventing to remove, prevent in the cultivation in-process, bump damage appears because of removing the cell, the bump and spacing frame 207 looks adaptation of gasbag post 206, the bottom of bull stick post 205 is provided with the rubber sleeve, the rubber sleeve of rack 205 is the sealed state with the rubber sleeve 206, the rack 208 is provided with the inside of rack 208, the inside life of rack is guaranteed, the rack is connected with the rack 209.
An immune cell CAR-T culture method with high cell survival rate comprises the following steps:
s1: preparation process of self-adding nutrient
When the cells are cultivated in the cultivation bin 302, when more oxygen and carbon dioxide are consumed, the carbon dioxide is used for adjusting the internal PH, constant PH is regulated according to the requirements of the cultivated cells, most of PH is cultivated under the condition of PH7.2-7.4, the PH change reacts with the reagent in the reaction tube 303 to form an electric signal which is transmitted to the electric signal receiver, then the electric signal receiver is instructed to increase the internal air pressure of the air bag block 306 to form an air pressure difference, and the air pressure difference drives the deflection rod 305 to rotate.
S2: nutrient addition step
Secondly, the deflection rod 305 deflects the outer side of the rotary valve 4 by squeezing and adjusting the outer side of the rotary valve 4 by a fixed distance, the deflection angle is 0-30 degrees, the rotary valve 4 is adjusted to open the switch of the internal air pressure, and the rotary valve 4 is adjusted to drive the sliding air bag 402 to be squeezed by sliding friction force between the rotary valve 4 and the sliding air bag 402, so that the air pressure drives the internal nutrient to be transferred into the cultivation cabin 302 through the transfer tube 401.
S3: alarm step of cultivation
If the cells undergo partial death or abnormal conditions in the cultivation process, the PH condition in the cultivation cabin 302 is greatly changed and is higher than PH7.6, so that the reaction with the reagent in the reaction tube 303 is stronger, an electric signal is formed and transmitted to the electric signal receiver, the electric signal receiver is instructed to drive the air bag block 306 to deflect the deflection rod 305 to 30-60 degrees, the curved clamping plate 304 limits the moving distance of the electric signal receiver, the deflection rod 305 drives the adjusting rotary valve 4 to rotate to an alarm area, and the equipment gives an alarm.
S4: emergency observation step
Excessive deflection of the adjusting rotary valve 4 drives the gear rack 205 to move backwards, the deflection angle of the gear rack 209 and the supporting frame 208 is limited, the gear rack 205 drives the auxiliary block 201 to move outwards, the auxiliary block 201 drives the auxiliary clamping block 203 to move outwards through the rotating rod column 202, the auxiliary clamping block 203 drives the sliding block 204 to move outwards, and then a worker can rapidly check specific conditions of internal cells through the observation window 101.
Working principle: when cells are cultivated in the cultivation bin 302, when more oxygen and carbon dioxide are consumed, the carbon dioxide is used for adjusting the internal PH, constant PH is regulated according to the requirements of cultivated cells, most PH is cultivated under the condition of PH7.2-7.4, the change of PH reacts with the reagent in the reaction tube 303, electric signals are transmitted to the electric signal receiver, the electric signals enable the internal air pressure of the air bag block 306 to be increased through instructions, air pressure difference is formed, the air pressure difference drives the deflection rod 305 to rotate, the deflection rod 305 is adjusted to deflect by a fixed distance through extrusion, the deflection angle is 0-30 degrees, the rotation valve 4 is adjusted to open the switch of the internal air pressure, the sliding air bag 402 is driven by sliding friction force between the rotation valve 4 and the sliding air bag 402 to be extruded, the air pressure drives the internal nutrient to be transmitted to the inside the cultivation bin 302 through the transmission tube 401, and therefore automatic nutrient conveying is achieved, and workers do not need to pay attention to specific cultivation conditions at any time, and the whole cultivation conditions are relatively large.
If the cells are in a biased death or abnormal condition in the cultivation process, the PH condition inside the cultivation bin 302 is greatly changed and higher than PH7.6, so that the reaction with the reagent inside the reaction tube 303 is stronger, an electric signal is formed and transmitted to the electric signal receiver, the electric signal receiver enables the airbag block 306 to drive the bias rod 305 to deflect to 30-60 degrees through the instruction, the curved clamping plate 304 limits the moving distance of the electric signal receiver, the bias rod 305 drives the regulating rotary valve 4 to rotate to an alarm area, the equipment sends an alarm, the excessive deflection of the regulating rotary valve 4 drives the gear rack 205 to move backwards, the gear rack 209 and the supporting frame 208 limit the deflecting angle of the gear rack, the gear rack 205 drives the auxiliary block 201 to move outwards, the auxiliary block 201 drives the auxiliary clamping block 203 to move outwards through the rotating rod column 202, and the auxiliary clamping block 203 drives the sliding block 204 to move outwards, so that staff can rapidly check the specific conditions of the cells inside through the observation window 101, and thus the survival rate of the cells is observed at the moment, and accidental cell death is avoided.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. Immune cell CAR-T culture apparatus that cell survival rate is high, including equipment frame (1), cultivate frame (3), adjustment rotary valve (4), its characterized in that: the device comprises an equipment outer frame (1) and is characterized in that the equipment outer frame (1) comprises an observation window (101), the cultivation frame (3) comprises a cultivation inner chamber (301), the cultivation frame (3) is arranged on the inner side of the equipment outer frame (1), a cultivation bin (302) for cultivating cells is arranged on the inner side of the cultivation inner chamber (301), the observation window (101) is arranged on the top end of a reaction tube (303), a reaction tube (303) for sensing the concentration of carbon dioxide in the interior is movably connected on the inner side of the cultivation bin (302), a transmission tube (401) for receiving nutrients is movably connected on one side, far away from the reaction tube (303), of the cultivation bin (302), an air bag block (306) for transmitting the internal air pressure is movably connected on the outer side of the reaction tube (303), a central rotating wheel (307) is movably connected on the outer side of the air bag block (306), curved clamping plates (304) are arranged on two sides of the deflection rod (305), an adjusting rotating valve (4) comprises a transmission tube (401), a sliding valve (402) is movably connected on the outer side of the transmission tube (401), and the adjusting valve (3) is arranged on the bottom end of the air bag.
The adjustable rotary valve is characterized in that the adjustable rotary valve (4) is internally provided with a staged latch, the staged latch of the adjustable rotary valve (4) and the sliding air bag (402) are arranged in an extrudable state, the staged latch of the adjustable rotary valve (4) is arranged in a deflection state under the pushing of the deflection rod (305), and the deflection distance is limited.
2. The immunocyte CAR-T culture apparatus having a high cell survival rate according to claim 1, wherein: the cell-observing device is characterized in that two sides of the outer frame (1) of the device are movably connected with a distance-adjusting frame (2) which is convenient for observing cells, a sealing ring is arranged on the outer side of the distance-adjusting frame (2), and the sealing ring of the distance-adjusting frame (2) is arranged in a sealing state with the outer frame (1) of the device.
3. The immunocyte CAR-T culture apparatus having a high cell survival rate according to claim 2, wherein: the distance-adjusting frame (2) comprises an auxiliary clamping block (203), a rotating rod column (202) used for deflecting is movably connected to the outer side of the auxiliary clamping block (203), and an auxiliary block (201) is movably connected to the outer side of the rotating rod column (202).
4. A high cell viability immune cell CAR-T culture apparatus according to claim 3, wherein: the two sides of the auxiliary block (201) are movably connected with gear racks (205) which are convenient to pull, the outer sides of the gear racks (205) are provided with limiting air bag columns (206), and two sides of each air bag column (206) are provided with limiting frames (207) which are used for preventing movement.
5. The high cell viability immune cell CAR-T culturing apparatus of claim 4 wherein: the number of the cultivation bins (302) is multiple, the cultivation bins (302) are arranged in a circular array with respect to the central rotating wheel (307), sealing blocks are arranged at the top ends of the cultivation bins (302), and the inside of the cultivation bins (302) is a sterile environment.
6. The high cell viability immune cell CAR-T culturing apparatus of claim 5 wherein: the utility model discloses a high-speed air bag with the structure, including gasbag post (206), gear rack (205), support frame (208) and support frame (208), the lug has been seted up to both sides of gasbag post (206), the lug and spacing (207) looks adaptation of gasbag post (206), the bottom of gear rack (205) is provided with the rubber sleeve, the rubber sleeve of gear rack (205) is sealed state setting with gasbag post (206), the bottom swing joint of gear rack (205) has support frame (208), the top swing joint of support frame (208) has commentaries on classics gear rack (209).
7. The high cell viability immune cell CAR-T culturing apparatus of claim 6 wherein: the number of the airbag blocks (306) and the deflection rods (305) is multiple, and the airbag blocks (306) and the deflection rods (305) are arranged in a circular ring shape relative to the central rotating wheel (307).
8. The cultivation method of an immunocyte CAR-T cultivation apparatus having a high cell survival rate according to claim 7, comprising the steps of:
s1: preparation process of self-adding nutrient
Firstly, when cells are cultivated in a cultivation bin (302), more oxygen is consumed and carbon dioxide is exhaled, the carbon dioxide reacts with reagents such as limestone in a reaction tube (303) to cause exothermic reaction, and at the moment, the internal air pressure of an air bag block (306) is reduced to form air pressure difference, and the air pressure difference drives a deflection rod (305) to rotate;
s2: nutrient addition step
Secondly, the deflection rod (305) deflects the outer side of the rotary valve (4) by extruding and adjusting the fixed distance, the deflection angle is 0-30 degrees, the rotary valve (4) is adjusted to open a switch of internal air pressure, the rotary valve (4) is adjusted to drive the sliding air bag (402) to be extruded through sliding friction force between the rotary valve and the sliding air bag (402), and then the air pressure drives internal nutrient to be transmitted into the cultivating bin (302) through the transmission pipe (401);
s3: alarm step of cultivation
If the cells are in partial death or abnormal conditions in the cultivation process, the mixing ratio of oxygen and carbon dioxide in the cultivation bin (302) is problematic, the pressure in the cultivation bin (302) is increased by excessive oxygen and nutrient which is not consumed, the internal pressure drives a deflection rod (305) to deflect to 30-60 degrees through an air bag block (306), a curved clamping plate (304) limits the moving distance of the cells, and the deflection rod (305) drives an adjusting rotary valve (4) to rotate to an alarm area, so that the equipment gives an alarm;
s4: emergency observation step
Excessive deflection of the adjusting rotary valve (4) drives the gear rack (205) to move backwards, the gear rack (209) and the supporting frame (208) limit the deflection angle of the gear rack, the gear rack (205) drives the auxiliary block (201) to move outwards, the auxiliary block (201) drives the auxiliary clamping block (203) to move outwards through the rotating rod column (202), the auxiliary clamping block (203) drives the sliding block (204) to move outwards, and then a worker can rapidly check the specific condition of internal cells through the observation window (101).
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| ITPI20040046A1 (en) * | 2004-06-18 | 2004-09-18 | Univ Pisa | BIOREACTOR FOR THE STUDY OF THE EFFECTS ON THE CELL ACTIVITIES OF IMPOSED STIMULES |
| CN111518695A (en) * | 2020-05-13 | 2020-08-11 | 易士力 | Biological cell culture dish |
| WO2020212384A1 (en) * | 2019-04-17 | 2020-10-22 | Hektros S.r.l. | Incubator for biological material |
| CN213877245U (en) * | 2020-12-15 | 2021-08-03 | 江苏万略医药科技有限公司 | In-vitro simulation device for pharmacokinetics |
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| US11285471B2 (en) * | 2017-05-30 | 2022-03-29 | Nextern Innovation, Llc | Portable controlled micro-environment device for modular bio-processing |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ITPI20040046A1 (en) * | 2004-06-18 | 2004-09-18 | Univ Pisa | BIOREACTOR FOR THE STUDY OF THE EFFECTS ON THE CELL ACTIVITIES OF IMPOSED STIMULES |
| WO2020212384A1 (en) * | 2019-04-17 | 2020-10-22 | Hektros S.r.l. | Incubator for biological material |
| CN111518695A (en) * | 2020-05-13 | 2020-08-11 | 易士力 | Biological cell culture dish |
| CN213877245U (en) * | 2020-12-15 | 2021-08-03 | 江苏万略医药科技有限公司 | In-vitro simulation device for pharmacokinetics |
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