CN114540193A - Breast cancer cell and T cell co-culture method for improving tumor killing capability of immune cells - Google Patents
Breast cancer cell and T cell co-culture method for improving tumor killing capability of immune cells Download PDFInfo
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- 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
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- 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
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- 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
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- C12N2502/00—Coculture with; Conditioned medium produced by
Abstract
The invention provides a CO-culture method of breast cancer cells and T cells for improving the capability of killing tumors by immune cells, which uses medical CO2The incubator is used for realizing; the medical CO2The incubator includes: the box, the main part of box is inside hollow structure, and the front end of box has the chamber door through inserting the post grafting, and the equal fixedly connected with in left end face four corners position of chamber door inserts the post. The invention solves the problem that the whole environment in the incubator is influenced when the incubator is taken out or put into the incubator to work in the prior art, and further the influence on the whole environment is influencedThe problem of the cultivation efficiency of cell, the partition control cover through guide block sliding connection on cultivateing the dish can relieve the closure to cultivateing the dish under magnetic block B's repulsion, and this design can realize that automatic completion under the electrically-controlled driven prerequisite of non-is opened and close confined structure to the automation of cultivateing the dish under the chamber door is opened and the non-state of opening, and then reaches the purpose of the inside culture of protection box.
Description
Technical Field
The invention relates to the technical field of medicine, in particular to a breast cancer cell and T cell co-culture method for improving the tumor killing capacity of immune cells.
Background
Breast cancer is considered to be an immune "cold" tumor, and only a small fraction of patients have an effective treatment with existing immune checkpoint inhibitors, with poor response rates, and therefore there is a critical need to explore new effective immunotherapeutic regimens.
In the prior application CN202210012502.0 by this research team, a new use of gamma interferon-induced lysosomal thiol reductase GILT in enhancing T cell-mediated anti-tumor immune responses was proposed. The results indicate that high expression of GILT can significantly increase the level of killer T cells in tumor tissues.
However, the above technical solutions do not relate to the specific co-culture method and the apparatus used in the method, and therefore, the present application proposes a co-culture method of breast cancer cells and T cells for improving the ability of immune cells to kill tumor.
With present incubator, current incubator is opening the back with the chamber door when cultivateing the work and cultivates the dish and mostly directly expose among the outside air for it can cause the influence to the inside holistic environment of incubator taking out or putting into the during operation, and then can influence its cultivation efficiency to the cell, and when cultivateing the work, lack the work of taking out and placing under the effectual reduction contact state, and then there is the limitation.
Disclosure of Invention
The invention provides a co-culture method of breast cancer cells and T cells for improving the capability of killing tumors by immune cells, and the co-culture method is realized by usingMedical CO2The incubator is used for realizing; the medical CO2The incubator includes: the box, the main part of box is inside hollow structure, and the front end of box has the chamber door through inserting the post grafting, the equal fixedly connected with in left end face four corners position of chamber door inserts the post, the chamber door has constituteed bearing structure jointly with inserting the post, it has seted up two logical grooves to be linear array in the inside of chamber door, and the logical inslot portion of seting up in the chamber door all articulates there is the apron, fixedly connected with guide arm on the face of the top of chamber door, the guide arm is equipped with two altogether, and two guide arms are linear array fixed connection in the face of the top of chamber door position, and keep away from one side of chamber door in two guide arms still fixedly connected with contact block, the contact block is the metal material, and the contact slot has been seted up on the face of contact block, contact slot and proximity switch phase-match are seted up to contact block bottom face.
Optionally, a partition plate is fixedly connected to the bottom end of the guide shaft, a culture tray is fixedly connected to the bottom end face of the partition plate, through grooves are formed in the left side and the right side of the culture tray, transverse grooves are formed in the front side and the rear side of the top end face of the culture tray, the transverse grooves formed in the top end of the culture tray are matched with the guide blocks, and top plates are fixedly connected to the left side face and the right side face of the partition plate.
Optionally, one side of the top plate, which is far away from the partition plate, is inserted into the partition control cover, the temperature controller is contained in the partition control cover, a guide block is fixedly connected to the bottom end face of the partition control cover, the partition control cover is connected onto the culture tray through the guide block in a sliding manner, folding plates are fixedly connected to the front side face and the rear side face of the partition control cover, and the partition control cover, the top plate and the folding plates form a closed structure.
Optionally, one side fixedly connected with fan of bellows, the fan is used for air-out in to bellows, is that the subtend articulates at the inboard of box and chamber door has support A, has still pegged graft at support A's the other end and has had support B, keeps away from support A's one end in support B and still is connected with the mount pad, at the inboard fixedly connected with tray of two mount pads, the inside of tray is rectangular array and has seted up logical groove, and the tray matches with the bottom opening of cultivateing the dish.
Optionally, the proximity switch is provided with two positions, the two proximity switches are respectively and fixedly connected to the left side and the right side of the top end face of the box body, the fan is in a power-on state when the proximity switch is in contact with the contact block, the top end face of the box body is fixedly connected with the magnetic blocks a, the magnetic blocks a are provided with two positions, and the two magnetic blocks a are fixedly connected to the top end face of the box body in a linear array manner.
Optionally, the partition control cover is made of a magnetic material, a magnetic block B is fixedly connected to the outer side of the box body, when the magnetic block B is overlapped with the partition control cover, the partition control cover is in a state of being in contact with the partition plate, a bottom plate is fixedly connected to the inner portion of the box body and used for sealing a bottom opening of the culture tray, and the culture tray and the bottom plate jointly form a bearing structure.
Optionally, the magnetic block a is matched with the contact block, a guide shaft is rotatably connected inside the box body, jacks are formed in the top end face of the guide shaft in an annular array, an adjusting disc is mounted at the top end of the guide shaft through a spring, guide pillars are arranged in the bottom end face of the adjusting disc in an annular array, and the guide pillars fixedly connected to the bottom end face of the adjusting disc are matched with the jacks formed in the top end of the guide shaft.
Optionally, one side of the bottom plate is fixedly connected with an air box, the air box is located under the culture tray, a through groove is formed in the air box, the through groove formed in the air box is matched with the groove formed in the bottom end of the culture tray, through longitudinal grooves are formed in the left side and the right side of the air box, and the longitudinal grooves formed in the left side and the right side of the air box are matched with the support A and the support B.
Advantageous effects
Be provided with bottom plate and bellows in the bottom side of cultivateing the dish, the bottom plate can regard as bearing and the enclosed construction to cultivateing the dish and use, and the setting of bellows not only can regard as the semi-enclosed passageway when passing through to the tray, and open the back when the chamber door, break away from through contact piece and proximity switch can also open the fan of installing in the bellows outside, and can be more comprehensive carry out more thoroughly and efficient wind to the tray of bellows inside and the article that inside held and drench the dust removal operation towards the inside of bellows through the fan after carrying out the air-out, and then reach the design that the article that the protection box inside held do not receive external environment influence, avoided suffering from the problem that the normal cultivation that leads to the fact the influence easily to the culture because of the environment interference of the box inside.
When the contact block is located at the position right above the magnetic block A, the contact block can be synchronously located at the position right below the adjusting disc, the adjusting disc arranged on the top side of the guide shaft is upwards jacked up through the protrusion of the contact block, the adjusting disc and the guide shaft are in a state of being separated from the non-transmission state, the position of the culture disc arranged in the box body cannot be adjusted by manually rotating the adjusting disc, the device can be more stable when being used for placing or taking out articles, inconvenience caused by mistaken touch is effectively prevented, and the design can be used for automatically and effectively avoiding that the device is easily damaged or the articles required to be stored or taken out are easily damaged and unnecessary loss is caused when the device is in the opening state of the box door if the position of the culture disc is carelessly adjusted.
The partition control cover connected on the culture tray in a sliding way can close the culture tray through the popping of the folding plate due to the loss of the limit of the magnetic block B, so that when the box door is in an open state, the partition control cover can close the culture tray to ensure that the cultured objects in the culture tray cannot be influenced by the interference of the external environment when the box door is in the open state, thereby achieving the more practical purpose, and when the culture tray is matched with the position of the magnetic block B fixedly connected on the outer side of the box body through the rotation of the adjusting plate, the partition control cover connected on the culture tray in a sliding way can remove the sealing of the culture tray under the repulsion action of the magnetic block B, the design can realize the automatic opening and closing closed structure of the culture tray under the opening and non-opening states of the box door under the premise of non-electrically controlled driving, thereby achieving the purpose of protecting the culture in the box body.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings of the embodiments will be briefly described below.
The drawings in the following description relate to some embodiments of the invention only and are not intended to limit the invention.
In the drawings:
FIG. 1 is a schematic diagram showing a front side view of an incubator according to an embodiment of the present invention in a partially constructed half-section and disassembled state;
FIG. 2 is a schematic view showing a bottom side view of the incubator according to an embodiment of the present invention in a partially constructed half-section and disassembled state;
FIG. 3 is a schematic view showing an assembled structure of an incubator according to an embodiment of the present invention;
FIG. 4 is a schematic diagram showing a box-to-magnet block B display structure of an incubator according to an embodiment of the invention;
FIG. 5 is a schematic diagram illustrating a guide shaft to guide block display structure of an incubator, according to an embodiment of the invention;
FIG. 6 shows a schematic view of a floor to mount display structure of an incubator, according to an embodiment of the invention;
FIG. 7 shows a schematic view of an enlarged structure at A in FIG. 2 of an incubator according to an embodiment of the invention;
FIG. 8 shows a schematic view of an enlarged structure at B in FIG. 2 of an incubator according to an embodiment of the invention;
list of reference numerals
1. A box body; 2. a box door; 3. a cover plate; 4. inserting a column; 5. a guide bar; 6. a contact block; 7. a proximity switch; 8. a magnetic block A; 9. a guide shaft; 10. an adjusting disk; 11. a guide post; 12. a culture tray; 13. a partition plate; 14. a zone control housing; 15. a top plate; 16. folding the board; 17. a guide block; 18. a base plate; 19. an air box; 20. a fan; 21. a bracket A; 22. a bracket B; 23. a tray; 24. a mounting seat; 25. and a magnetic block B.
Detailed Description
In order to make the objects, aspects and advantages of the embodiments of the present invention more apparent, the embodiments of the present invention will be described in detail with reference to the accompanying drawings. Unless otherwise indicated, terms used herein have the ordinary meaning in the art. Like reference symbols in the various drawings indicate like elements.
Example (b): please refer to fig. 1 to 8:
the invention provides a CO-culture method of breast cancer cells and T cells for improving the capability of killing tumors by immune cells, which uses medical CO2The incubator is used for realizing; the medical CO2The incubator includes: the box body 1, the main body of the box body 1 is an internal hollow structure, the front end of the box body 1 is inserted with a box door 2 through an inserting column 4, four corners of the left end surface of the box door 2 are fixedly connected with the inserting column 4, the box door 2 and the inserting column 4 jointly form a supporting structure, two through grooves are arranged in the box door 2 in a linear array manner, cover plates 3 are hinged inside the through grooves arranged in the box door 2, guide rods 5 are fixedly connected to the top end surface of the box door 2, the guide rods 5 are arranged in two positions, the two guide rods 5 are fixedly connected to the top end surface of the box door 2 in a linear array manner, one side of the two guide rods 5 far away from the bottom end surface of the box door 2 is also fixedly connected with a contact block 6, the contact block 6 is made of metal material, a contact groove is arranged on the bottom end surface of the contact block 6, the contact groove arranged on the surface of the contact block 6 is matched with a proximity switch 7, one side of a fan 20 is fixedly connected with the air box 19, the fan 20 is used for blowing air into the bellows 19, a support A21 is hinged on the inner sides of the box body 1 and the box door 2 in an opposite direction, a support B22 is further inserted at the other end of the support A21, one end of the support B22, which is far away from the support A21, is further connected with the mounting seat 24, a tray 23 is fixedly connected with the inner sides of the two mounting seats 24, a through groove is formed in the tray 23 in a rectangular array, the tray 23 is matched with the bottom end opening of the culture tray 12, when the contact block 6 is positioned at the position right above the magnetic block A8, the contact block 6 can be synchronously positioned at the position right below the adjusting tray 10, the adjusting tray 10 arranged at the top side of the guide shaft 9 is jacked upwards through the protrusion of the contact block 6, the adjusting tray 10 and the guide shaft 9 are in a state of being separated from being incapable of transmission, the adjusting tray 10 cannot be manually rotated to adjust the position of the culture tray 12 arranged in the box body 1, and articles can be put in or taken out more stably when the device is used, and inconvenience caused by mistaken touch is effectively prevented.
In addition, according to the embodiment of the present invention, as shown in fig. 1 to fig. 3, the proximity switch 7 has two positions, and the two proximity switches 7 are respectively and fixedly connected to the left and right side positions on the top end surface of the case 1, and the fan 20 is in the power-on state in the state that the proximity switch 7 is in contact with the contact block 6, the top end surface of the case 1 is fixedly connected with the magnetic block A8, the magnetic block A8 has two positions, and the two magnetic blocks A8 are fixedly connected to the top end surface of the case 1 in a linear array, the magnetic block A8 is matched with the contact block 6, the inside of the case 1 is rotatably connected with the guide shaft 9, the top end surface of the guide shaft 9 is provided with the insertion holes in an annular array, the top end of the guide shaft 9 is provided with the adjustment disc 10 through the spring, the guide posts 11 are arranged in an annular array on the bottom end surface of the adjustment disc 10, and the guide posts 11 fixedly connected to the bottom end surface of the adjustment disc 10 are matched with the insertion holes provided on the top end of the guide shaft 9, the box door 2 is pulled outwards through manual operation, the contact block 6 arranged on one side of the guide rod 5 can be separated from the proximity switch 7 fixedly connected to the box body 1, the fan 20 arranged on the outer side of the bellows 19 is in an opening state, then the box door 2 is continuously opened outwards, the contact block 6 arranged on one side of the guide rod 5 is located right above the magnetic block A8 fixedly connected to the top end of the box body 1, and the box door 2 can be effectively limited after being opened through the adsorption of the magnetic block A8 on the contact block 6.
In addition, according to the embodiment of the present invention, as shown in fig. 1 to 6, a partition plate 13 is fixedly connected to the bottom end of the guide shaft 9, a culture tray 12 is fixedly connected to the bottom end surface of the partition plate 13, through grooves are formed on both left and right sides of the culture tray 12, transverse grooves are formed on both front and rear sides of the top end surface of the culture tray 12, the transverse grooves formed on the top end of the culture tray 12 are matched with guide blocks 17, top plates 15 are fixedly connected to both left and right sides of the partition plate 13, one side of the top plate 15 away from the partition plate 13 is inserted into a partition control cover 14, a temperature controller is contained in the partition control cover 14, the guide blocks 17 are fixedly connected to the bottom end surface of the partition control cover 14, the partition control cover 14 is slidably connected to the culture tray 12 through the guide blocks 17, folding plates 16 are fixedly connected to both front and rear sides of the partition control cover 14, and the partition control cover 14, the top plate 15 and the folding plates 16 constitute a closed structure, when the culture tray 12 is matched with the position of the magnetic block B25 fixedly connected with the outer side of the box body 1 through the rotation of the adjusting tray 10, the partition control cover 14 which is connected onto the culture tray 12 in a sliding mode through the guide block 17 can release the sealing of the culture tray 12 under the repulsion action of the magnetic block B25, and the aim of automatic adjustment is achieved.
In addition, according to the embodiment of the present invention, as shown in fig. 1 to 8, the partitioned control cover 14 is made of a magnetic material, a magnetic block B25 is fixedly connected with the outer side of the box body 1, when the magnetic block B25 is superposed with the position of the subarea control cover 14, the subarea control cover 14 is in a contact state with the clapboard 13, a bottom plate 18 is fixedly connected in the box body 1, the bottom plate 18 is used for sealing the bottom end opening of the culture tray 12, and the culture tray 12 and the bottom plate 18 form a bearing structure together, one side of the bottom plate 18 is fixedly connected with an air box 19, the air box 19 is positioned right below the culture tray 12, and a through groove is arranged inside the air box 19, the through groove arranged in the air box 19 is matched with the groove at the bottom end of the culture tray 12, the left side and the right side of the air box 19 are both provided with through longitudinal grooves, and the longitudinal grooves arranged on the left side and the right side of the air box 19 are matched with the support A21 and the support B22.
The co-culture method comprises the following steps:
when the culture tray 12 mounted on the bottom side of the guide shaft 9 is left empty for storing articles, when the box door 2 is mounted at the front position of the box body 1 and the box body 1 is in a closed state, the adjusting tray 10 mounted on the top side of the guide shaft 9 is pressed downwards, so that the guide post 11 fixedly connected with the bottom side of the adjusting tray 10 is inserted into the insertion hole formed in the top end of the guide shaft 9, the adjusting tray 10 is manually rotated, the culture tray 12 mounted on the bottom side of the partition plate 13 is driven by the guide shaft 9 to rotate, and one side of the culture tray 12 provided with the through groove faces to one side of the box door 2;
then the box door 2 is pulled outwards manually, firstly the contact block 6 arranged on one side of the guide rod 5 can be separated from the proximity switch 7 fixedly connected on the box body 1, at the moment, the fan 20 arranged on the outer side of the air box 19 is in an opening state, then the box door 2 is continuously opened outwards, so that the contact block 6 arranged on one side of the guide rod 5 is positioned right above the magnetic block A8 fixedly connected at the top end of the box body 1, and the box door 2 can be effectively limited after being opened by the adsorption of the magnetic block A8 on the contact block 6;
when the contact block 6 is positioned right above the magnetic block A8, the contact block is synchronously positioned right below the adjusting disc 10, the adjusting disc 10 arranged on the top side of the guide shaft 9 is jacked upwards through the bulge of the contact block 6, so that the adjusting disc 10 and the guide shaft 9 are in a state of being separated from each other and incapable of transmission, the position of the culture disc 12 arranged in the box body 1 cannot be adjusted by manually rotating the adjusting disc 10, articles can be more stably put in or taken out when the device is used, and inconvenience caused by mistaken touch is effectively prevented;
at this time, the tray 23 slidably connected to the inner side of the bellows 19 moves downward along the bellows 19 under the traction of the support a21 and the support B22, when the tray 23 moves downward and is completely separated from the bellows 19, the articles can be placed on the tray 23 for carrying, and then the door 2 and the box body 1 are manually closed, the articles can be automatically guided to the inner position of the cultivation tray 12 for containing by the carrying of the tray 23, and in the process that the tray 23 ascends or descends along the bellows 19, the fan 20 installed on the outer side of the bellows 19 blows air to the inner part of the bellows 19 to perform air shower and dust removal operations on the tray 23 and the articles placed above the tray 23, so as to achieve the purpose of reducing the influence of the external environment on the environment inside of the box body 1 as much as possible;
when the narrower side of the incubator tray 12 faces the direction of the door 2, the partition control cover 14 slidably connected to the incubator tray 12 through the guide block 17 can close the incubator tray 12 by popping up the folding plate 16 due to the loss of the position limitation of the magnetic block B25, so that when the door 2 is in an open state, the partition control cover 14 can close the incubator tray 12 to ensure that the incubation objects in the incubator tray 12 can not be influenced by the interference of the external environment when the door 2 is in an open state, thereby achieving a more practical purpose;
when the position of the magnetic block B25 fixedly connected with the outer side of the box body 1 is matched with the culture tray 12 through the rotation of the adjusting disk 10, the partition control cover 14 which is connected onto the culture tray 12 in a sliding mode through the guide block 17 can release the sealing of the culture tray 12 under the repulsion action of the magnetic block B25, and the aim of automatic adjustment is achieved.
Except for the specific use of medical CO2Apart from the incubator step, the complete process of the co-cultivation method can be performed with reference to CN202210012502.0, i.e. comprising the following steps:
experimental materials:
cell lines: activating T cells, human breast cancer cell strains MDA-MB-231 and BT-549
Reagent consumables: cytotoxic Lactate Dehydrogenase (LDH) assay kit, T cell culture medium, cell fixative solution, crystal violet solution, and cell culture plate
The instrument comprises the following steps: cell culture case, superclean bench, enzyme-labeling instrument
The experimental method comprises the following steps:
LDH experiment:
(1) 100. mu.L of breast cancer cells (cell number: 5000) were pipetted into a 96-well plate and pre-cultured overnight at 37 ℃ in a 5% CO2 incubator;
(2) to the breast cancer cells, 100. mu.L of a medium containing 20 ten thousand T cells was added, and the culture was continued at 37 ℃ in a CO2 incubator.
(3) Adding 10 μ L lysine Buffer into CO-cultured breast cancer cells, and incubating at 37 deg.C in 5% CO2 incubator for 30 min;
(4) adding 100 mu L of Working Solution into each hole, and incubating for 10min under the conditions of dark and room temperature;
(5) add 50. mu.L of Stop Solution to each well and immediately measure absorbance at 490nM wavelength using a microplate reader.
(6) The calculation method comprises the following steps:
the absorbance of "sample well-sample Blank well", "high control well-high control Blank well", "low control well-background Blank well" was calculated, and then the average value of n-3 was calculated.
The cell damage rate was calculated according to the following formula:
cell damage rate (%) [ (a-C)/(B-C) ] × 100
Absorbance of sample (sample well-sample Blank well)
High control Absorbance (high control well-high control Blank well)
C absorbance of low control (low control wells-background Blank wells).
As a result: after co-culture of the breast cancer cells and the T cells, the killing rate of the T cells to GILT over-expressed breast cancer cells is obviously higher than that of a control group.
Crystal violet staining experiment:
(1) inoculating 500 μ L of breast cancer cells (cell number: 1 ten thousand) to a 24-well plate, and pre-culturing overnight in a 5% CO2 incubator at 37 deg.C;
(2) adding 2mL of a culture medium containing 40 ten thousand T cells into the breast cancer cells, and continuously culturing for 5 days in a 5% CO2 incubator at 37 ℃;
(3) the culture medium is sucked and removed, PBS is used for washing for 3 times, and cell is fixed for 30min by using cell fixing solution;
(4) and (3) staining the fixed cells for 30min by using a crystal violet solution, slowly washing away a staining solution by using running water, drying in air, and photographing and recording.
The experimental results are as follows:
as shown in fig. 3 and 4, the killing effect of T cells on GILT-highly expressed breast cancer cells was significantly increased compared to the control group after co-culture.
In addition to specific co-culture, the whole process of the application of gamma interferon-induced lysosomal thiol reductase GILT in enhancing T cell-mediated anti-tumor immune response can be performed with reference to CN202210012502.0, including model construction, T cell sorting and activation, GILT enhancing killing ability of T cells to tumor cells, high expression GILT significantly improving the level of infiltrating T cells in breast cancer tissues, and the like.
Finally, it should be noted that, when describing the positions of the components and the matching relationship therebetween, the present invention is usually illustrated by one/a pair of components, however, it should be understood by those skilled in the art that such positions, matching relationship, etc. are also applicable to other/other pairs of components.
The above description is intended to be illustrative of the present invention and not to limit the scope of the invention, which is defined by the claims appended hereto.
Claims (8)
1. A CO-culture method of breast cancer cells and T cells for improving the ability of immune cells to kill tumors, which is characterized in that the CO-culture method uses medical CO2The incubator is used for realizing; the medical CO2The incubator includes: the box body (1), the main body of the box body (1) is of an internal hollow structure, the front end of the box body (1) is inserted with a box door (2) through an inserting column (4), four corners of the left end face of the box door (2) are fixedly connected with the inserting column (4), the box door (2) and the inserting column (4) jointly form a supporting structure, two through grooves are formed in the box door (2) in a linear array manner, a cover plate (3) is hinged inside each through groove formed in the box door (2), a guide rod (5) is fixedly connected to the top end face of the box door (2), the guide rod (5) is provided with two positions, the two guide rods (5) are fixedly connected to the top end face of the box door (2) in a linear array manner, a contact block (6) is further fixedly connected to one side, far away from the box door (2), of the two guide rods (5), the contact block (6) is made of metal, and a contact groove is formed in the bottom end face of the contact block (6), the contact groove arranged on the bottom end surface of the contact block (6) is matched with the proximity switch (7).
2. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 1, wherein: the proximity switch (7) is provided with two positions, the two proximity switches (7) are respectively fixedly connected to the left side and the right side of the top end face of the box body (1), the fan (20) is in a power-on state under the state that the proximity switches (7) are in contact with the contact blocks (6), the top end face of the box body (1) is fixedly connected with magnetic blocks A (8), the magnetic blocks A (8) are provided with two positions, and the two magnetic blocks A (8) are fixedly connected to the top end face of the box body (1) in a linear array mode.
3. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 2, wherein: the magnetic block A (8) is matched with the contact block (6), a guide shaft (9) is rotatably connected inside the box body (1), jacks are formed in the top end face of the guide shaft (9) in an annular array mode, an adjusting disc (10) is installed on the top end of the guide shaft (9) through a spring, guide pillars (11) are arranged on the bottom end face of the adjusting disc (10) in an annular array mode, and the guide pillars (11) fixedly connected to the bottom end face of the adjusting disc (10) are matched with the jacks formed in the top end of the guide shaft (9).
4. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 3, wherein: bottom fixedly connected with baffle (13) of guide shaft (9), fixedly connected with cultivates dish (12) on the bottom face of baffle (13), cultivates the left and right sides of dish (12) and has all seted up logical groove, and cultivates the top face front and back both sides of dish (12) and has all seted up the transverse groove to cultivate the transverse groove and guide block (17) phase-match that the dish (12) top was seted up, at the equal fixedly connected with roof (15) of the left and right sides face of baffle (13).
5. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 4, wherein: one side of the top plate (15) far away from the partition plate (13) is connected with a partition control cover (14) in an inserting mode, a temperature controller is arranged inside the partition control cover (14), a guide block (17) is fixedly connected to the bottom end face of the partition control cover (14), the partition control cover (14) is connected onto the culture disc (12) in a sliding mode through the guide block (17), folding plates (16) are fixedly connected to the front side face and the rear side face of the partition control cover (14), and the partition control cover (14), the top plate (15) and the folding plates (16) form a closed structure.
6. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 5, wherein: the partition control cover (14) is made of magnetic materials, a magnetic block B (25) is fixedly connected to the outer side of the box body (1), when the magnetic block B (25) is overlapped with the partition control cover (14), the partition control cover (14) is in a contact state with the partition plate (13), a bottom plate (18) is fixedly connected to the inner portion of the box body (1), the bottom plate (18) is used for sealing a bottom end opening of the culture disc (12), and the culture disc (12) and the bottom plate (18) jointly form a bearing structure.
7. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 6, wherein: one side fixedly connected with bellows (19) of bottom plate (18), bellows (19) are located cultivates dish (12) and put under, and the inside of bellows (19) has seted up logical groove to logical groove that set up in bellows (19) and the bottom fluting phase-match of cultivateing dish (12), all set up the vertical groove that link up in the left and right sides of bellows (19), and the vertical groove that the left and right sides was seted up in bellows (19) and support A (21) and support B (22) phase-match.
8. The method of co-culturing breast cancer cells and T cells to enhance the ability of immune cells to kill tumors as defined in claim 7, wherein: one side fixedly connected with fan (20) of bellows (19), fan (20) are used for the air-out in bellows (19), it has support A (21) to be that the subtend articulates in the inboard of box (1) and chamber door (2), support B (22) have still been pegged graft to the other end at support A (21), the one end of keeping away from support A (21) in support B (22) still is connected with mount pad (24), inboard fixedly connected with tray (23) at two mount pads (24), the inside of tray (23) is rectangular array and has seted up logical groove, and tray (23) and the bottom opening phase-match of cultivateing dish (12).
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Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204607041U (en) * | 2015-03-18 | 2015-09-02 | 青岛银泰洁净设备有限公司 | A kind of novel air spraying type pass-through box |
| CN206127328U (en) * | 2016-09-21 | 2017-04-26 | 遵义医学院 | Antitumor cell culture device for immunity |
| CN207193300U (en) * | 2017-09-12 | 2018-04-06 | 苏州工业园区旭太生物工程有限公司 | A kind of cell culture medium drying system |
| EP3511719A1 (en) * | 2016-09-08 | 2019-07-17 | Hitachi High-Technologies Corporation | Automatic analysis device |
| CN210178143U (en) * | 2019-05-20 | 2020-03-24 | 浙江欣世纪建筑系统科技有限公司 | Sliding window capable of being automatically positioned |
| CN210309470U (en) * | 2019-04-28 | 2020-04-14 | 江苏欧泰电子工业集团有限公司 | Various steel sheet of rock wool production purifier |
| CN211771224U (en) * | 2019-12-25 | 2020-10-27 | 陕西泾河好邦食品有限公司 | Microbial cultivation case |
| CN212559296U (en) * | 2020-04-30 | 2021-02-19 | 重庆德庄农产品开发有限公司 | Raw material transfer box capable of automatically transferring |
| CN212925007U (en) * | 2020-08-04 | 2021-04-09 | 广州市土根旺生物科技有限公司 | Compound fungus crowd breeds culture apparatus |
| CN213001560U (en) * | 2020-09-01 | 2021-04-20 | 成都金田种苗有限公司 | Seedling growing greenhouse wind drenches degassing unit |
| CN214400525U (en) * | 2021-03-26 | 2021-10-15 | 安徽省点滴环境科技有限公司 | Novel environment detects cultivates with constant temperature device |
| CN214458059U (en) * | 2020-12-01 | 2021-10-22 | 上海润度生物科技有限公司 | Dustproof and moistureproof carbon dioxide incubator structure |
| CN214937463U (en) * | 2021-04-06 | 2021-11-30 | 矫文文 | A incubator for food detection |
| CN114306582A (en) * | 2022-01-07 | 2022-04-12 | 山东大学 | Application of gamma interferon-induced lysosome sulfhydryl reductase GILT in enhancing anti-tumor immune response |
-
2022
- 2022-03-14 CN CN202210244547.0A patent/CN114540193A/en active Pending
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN204607041U (en) * | 2015-03-18 | 2015-09-02 | 青岛银泰洁净设备有限公司 | A kind of novel air spraying type pass-through box |
| EP3511719A1 (en) * | 2016-09-08 | 2019-07-17 | Hitachi High-Technologies Corporation | Automatic analysis device |
| CN206127328U (en) * | 2016-09-21 | 2017-04-26 | 遵义医学院 | Antitumor cell culture device for immunity |
| CN207193300U (en) * | 2017-09-12 | 2018-04-06 | 苏州工业园区旭太生物工程有限公司 | A kind of cell culture medium drying system |
| CN210309470U (en) * | 2019-04-28 | 2020-04-14 | 江苏欧泰电子工业集团有限公司 | Various steel sheet of rock wool production purifier |
| CN210178143U (en) * | 2019-05-20 | 2020-03-24 | 浙江欣世纪建筑系统科技有限公司 | Sliding window capable of being automatically positioned |
| CN211771224U (en) * | 2019-12-25 | 2020-10-27 | 陕西泾河好邦食品有限公司 | Microbial cultivation case |
| CN212559296U (en) * | 2020-04-30 | 2021-02-19 | 重庆德庄农产品开发有限公司 | Raw material transfer box capable of automatically transferring |
| CN212925007U (en) * | 2020-08-04 | 2021-04-09 | 广州市土根旺生物科技有限公司 | Compound fungus crowd breeds culture apparatus |
| CN213001560U (en) * | 2020-09-01 | 2021-04-20 | 成都金田种苗有限公司 | Seedling growing greenhouse wind drenches degassing unit |
| CN214458059U (en) * | 2020-12-01 | 2021-10-22 | 上海润度生物科技有限公司 | Dustproof and moistureproof carbon dioxide incubator structure |
| CN214400525U (en) * | 2021-03-26 | 2021-10-15 | 安徽省点滴环境科技有限公司 | Novel environment detects cultivates with constant temperature device |
| CN214937463U (en) * | 2021-04-06 | 2021-11-30 | 矫文文 | A incubator for food detection |
| CN114306582A (en) * | 2022-01-07 | 2022-04-12 | 山东大学 | Application of gamma interferon-induced lysosome sulfhydryl reductase GILT in enhancing anti-tumor immune response |
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