CN215947322U

CN215947322U - Culture device for establishing stable transfection cell line of Cyr61 target

Info

Publication number: CN215947322U
Application number: CN202122378015.XU
Authority: CN
Inventors: 石卫东
Original assignee: Fudan University Shanghai Cancer Center
Current assignee: Fudan University Shanghai Cancer Center
Priority date: 2021-09-29
Filing date: 2021-09-29
Publication date: 2022-03-04
Anticipated expiration: 2031-09-29

Abstract

The utility model discloses a culture device for establishing a Cyr61 target stable transfection cell line, which comprises a containing component and a material supplementing component, wherein the containing component comprises a box body and a box door, the box door is hinged and installed on the side part of the box body, a culture barrel is installed inside the box body, the material supplementing component comprises a plurality of material guide pipes, a plurality of electromagnetic valves, a material mixing tank and a stirring piece, the plurality of material guide pipes are annularly arranged on the side part of the culture barrel in an array mode, the plurality of electromagnetic valves are respectively installed in the middle parts of the plurality of material guide pipes, the plurality of material guide pipes extend out of the upper part of the box body, the material mixing tank is installed on the upper parts of the plurality of material guide pipes, two material supplementing pipes are installed on the upper part of the material supplementing pipe, a cover plate is installed on the upper part of the material mixing tank, and the stirring piece is installed in the center of the upper part of the material mixing tank. According to the utility model, the material supplementing process can be realized without opening the box door, the interference of external bacteria to the culture medium is reduced, and the subsequent culture of a cell line is ensured.

Description

Culture device for establishing stable transfection cell line of Cyr61 target

Technical Field

The utility model relates to the technical field of cell line culture, in particular to a culture device for establishing a Cyr61 target stable transfection cell line.

Background

The prognosis of the liver metastasis of the pancreatic cancer is poor, so far, an effective treatment means is lacked, and the search for a new treatment target point has important clinical value. Project group early-stage projects prove that Cyr61 is regulated to regulate EMT to inhibit pancreatic cancer liver metastasis when clearing pancreas stasis, and in subsequent researches, people find that the pancreas stasis clearing formula can inhibit tumor-associated macrophage (TAM) infiltration and IL-6 secretion. Therefore, the hypothesis is proposed: the effect of regulating EMT to inhibit pancreatic cancer liver metastasis by Cyr61 in clearing pancreas accumulation is dependent on the participation of TAM in a tumor microenvironment, and Cyr61 can promote the growth and metastasis of pancreatic cancer cells by regulating and controlling a tumor-related macrophage-pancreatic cancer cell interaction feedback loop. In order to verify the hypothesis, the action mechanism of Cyr61 for regulating and controlling tumor-associated macrophage-pancreatic cancer cell interaction and promoting the growth and metastasis of pancreatic cancer cells is determined by means of transfection of liposome vectors, a pancreatic cancer cell and macrophage co-culture system and the like.

In the verification process, an in vitro experiment that Cyr61 regulates tumor-related macrophage-pancreatic cancer cell interaction to promote growth and transfer of pancreatic cancer cells needs to be carried out, and a Cyr61 stable transfection cell line needs to be established.

However, in the existing culture device, the box door needs to be opened manually, and G418 antibiotics and nutrients are supplemented to the box door in the period so as to maintain the culture condition, and bacteria are easy to enter in the process of opening the box door, so that the culture of the cell line is influenced.

SUMMERY OF THE UTILITY MODEL

The utility model aims to provide a culture device for establishing a Cyr61 target stable transfection cell line, so as to solve the problems in the background technology.

In order to achieve the purpose, the utility model provides the following technical scheme: a culture device for establishing a Cyr61 target stable transfection cell line comprises a containing component and a feeding component.

The accommodating assembly comprises a box body and a box door, the box door is hinged to the side portion of the box body, and a culture barrel is arranged inside the box body.

The feed supplement subassembly includes a plurality of passage pipes, a plurality of solenoid valves, compounding jar and stirring piece, and is a plurality of passage pipe circular array install in the lateral part of cultivateing the bucket, it is a plurality of the solenoid valve is installed respectively in a plurality of the middle part of passage pipe, a plurality of the passage pipe is followed the upper portion of box stretches out, the compounding jar is installed in a plurality of the upper portion of passage pipe, two feed supplement pipes are installed on the upper portion of compounding jar, the apron is installed on the upper portion of feed supplement pipe, the stirring piece install in the upper portion center of compounding jar.

In one embodiment of the present invention, an observation window is installed on a side wall of the box body, and a foot seat is connected to a lower wall of the box body.

In an embodiment of the present invention, the cross-section of the material guiding pipe is provided in a J shape, and the inner wall of the material guiding pipe is provided with a smooth surface.

In one embodiment of the utility model, a lamp holder is arranged on the inner side of the upper wall of the mixing tank, and an ultraviolet lamp is arranged on the lower part of the lamp holder.

In an embodiment of the utility model, the stirring member includes a driving motor, a supporting shaft and a plurality of stirring blades, the driving motor is mounted on the upper wall of the mixing tank, the supporting shaft is mounted at the output end of the driving motor, the supporting shaft extends into the inner side of the mixing tank, and the plurality of stirring blades are respectively mounted on the side portions of the supporting shaft.

In one embodiment of the utility model, the side part of the stirring blade is provided with a scraper, and the side wall of the scraper is attached to the inner wall of the mixing tank.

In summary, due to the adoption of the technology, the utility model has the beneficial effects that:

according to the utility model, the sterilized liquid culture medium and the G418 antibiotic are introduced into the mixing tank, the stirring is carried out through the operation of the stirring piece, so that the liquid culture medium and the G418 antibiotic are uniformly mixed, then the electromagnetic valve is opened, so that the mixed material enters the culture barrel through the material guide pipe, and the clone body is cultured in the culture barrel, thereby avoiding the situation that the box door is directly opened to supplement the material, and reducing the possibility of external bacterial pollution.

Drawings

FIG. 1 is a schematic front view of the present invention;

FIG. 2 is a schematic front sectional view of the present invention;

FIG. 3 is a schematic top view of a part of the structure of a culture tank according to the present invention;

FIG. 4 is a schematic sectional front view of the mixing bowl of the present invention.

In the figure: 100. a containment assembly; 110. a box body; 111. an observation window; 112. a foot seat; 120. a box door; 130. a culture bucket; 200. a feed supplement assembly; 210. a material guide pipe; 220. an electromagnetic valve; 230. a mixing tank; 231. a lamp socket; 232. an ultraviolet lamp; 240. a stirring member; 241. a drive motor; 242. a support shaft; 243. a stirring blade; 244. a squeegee; 250. a material supplementing pipe; 260. and (7) a cover plate.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the utility model, as claimed, but is merely representative of selected embodiments of the utility model. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

In the description of the present invention, it is to be understood that the terms indicating an orientation or positional relationship are based on the orientation or positional relationship shown in the drawings only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art from the specification.

Example 1

Referring to fig. 1 to 4, the present invention provides a culture device for establishing a Cyr61 target stable transfected cell line, comprising a holding assembly 100 and a feeding assembly 200, wherein the feeding assembly 200 is mounted on the holding assembly 100.

The accommodating component 100 comprises a box body 110 and a box door 120, the box door 120 is hinged to the side portion of the box body 110, a culture barrel 130 is installed in the box body 110 through bolts, when the accommodating component is specifically arranged, an observation window 111 is installed on the side wall of the box body 110, a foot base 112 is connected to the lower wall of the box body 110, the observation window 111 is convenient to observe the inside of the box body 110, the culture condition can be conveniently known, and the foot base 112 is convenient to stably support the box body 110.

It should be noted that the door 120 is sealed with the box body 110 after being closed, a temperature adjusting device for adjusting the temperature is installed inside the box body 110 to provide the temperature required for cultivation, and the box body 110 is provided with a sterilizing and ventilating device to provide the oxygen required for cultivation.

The feeding assembly 200 comprises a plurality of material guide pipes 210, a plurality of electromagnetic valves 220, a material mixing tank 230 and a stirring piece 240, wherein the plurality of material guide pipes 210 are arranged on the side part of the culture bucket 130 in an annular array manner, when the feeding assembly is specifically arranged, in order to facilitate the material to be cultured in the culture bucket 130, the sections of the material guide pipes 210 are arranged in a J shape, and the inner walls of the material guide pipes 210 are arranged to be smooth surfaces; the plurality of electromagnetic valves 220 are respectively arranged at the middle parts of the plurality of material guiding pipes 210, and in order to prevent the initial material from falling into the material guiding pipes 210 in an excessive amount in the adding process, the electromagnetic valves 220 are arranged at the positions adjacent to the material mixing tank 230 as much as possible; the material guiding pipes 210 extend out of the upper part of the box body 110, the material mixing tank 230 is arranged on the upper parts of the material guiding pipes 210, and when the material mixing tank is specifically arranged, in order to achieve a better disinfection effect, the interior of the material mixing tank 230 is disinfected, a lamp holder 231 is arranged on the inner side of the upper wall of the material mixing tank 230, and an ultraviolet lamp 232 is arranged on the lower part of the lamp holder 231; two feeding pipes 250 are installed on the upper portion of the mixing bowl 230, a cover plate 260 is threadedly installed on the upper portion of the feeding pipes 250, and a stirring member 240 is installed at the center of the upper portion of the mixing bowl 230.

In this embodiment, the stirring part 240 includes a driving motor 241, a supporting shaft 242 and a plurality of stirring blades 243, the driving motor 241 is installed on the upper wall of the mixing tank 230 through bolts, the supporting shaft 242 is installed at the output end of the driving motor 241, the supporting shaft 242 extends into the inner side of the mixing tank 230, the plurality of stirring blades 243 are respectively installed at the side portions of the supporting shaft 242, during specific setting, the side portions of the stirring blades 243 are provided with scraping plates 244 through screws, the side walls of the scraping plates 244 are attached to the inner wall of the mixing tank 230, during specific use, the supporting shaft 242 is driven to rotate through the operation of the driving motor 241, the rotation of the supporting shaft 242 drives the stirring blades 243 to rotate, materials entering the mixing tank 230 are mixed, and the scraping plates 244 are used for scraping off the materials hung on the wall, so as to achieve a good stirring effect.

The working principle is as follows: during the use, open apron 260, respectively through two material supplement pipes 250, let in liquid medium and the G418 antibiotic after the disinfection to the inside of compounding jar 230, stir through the operation of stirring piece 240, make liquid medium and G418 antibiotic misce bene, then open solenoid valve 220, make the material after mixing enter into the inside of culture vat 130 through passage 210, carry out the cultivation of the clone body in culture vat 130, avoided directly opening chamber door 120 and supply the material, the reduction takes place the possibility of outside bacterial contamination.

It should be noted that: the model specifications of the electromagnetic valve 220, the ultraviolet lamp 232 and the driving motor 241 need to be determined according to the actual specification of the device, and the specific model selection calculation method adopts the prior art in the field, so detailed description is omitted.

The power supply and the principle of the solenoid valve 220, the ultraviolet lamp 232, and the driving motor 241 will be apparent to those skilled in the art and will not be described in detail herein.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and equivalent alternatives or modifications according to the technical solution of the present invention and the inventive concept thereof should be covered by the scope of the present invention.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

Claims

1. A culture device for establishing a Cyr61 target stable transfection cell line, which is characterized by comprising:

the container comprises a containing assembly (100), wherein the containing assembly (100) comprises a box body (110) and a box door (120), the box door (120) is hinged to the side part of the box body (110), and a culture bucket (130) is installed inside the box body (110);

feed supplement subassembly (200), feed supplement subassembly (200) include a plurality of passage tubes (210), a plurality of solenoid valve (220), compounding jar (230) and stirring piece (240), and is a plurality of passage tube (210) annular array install in the lateral part of cultivateing bucket (130), it is a plurality of solenoid valve (220) are installed respectively in a plurality of the middle part of passage tube (210), it is a plurality of passage tube (210) are followed the upper portion of box (110) stretches out, install in a plurality of compounding jar (230) the upper portion of passage tube (210), two material supplementing pipes (250) are installed on the upper portion of compounding jar (230), apron (260) are installed on the upper portion of material supplementing pipe (250), stirring piece (240) install in the upper portion center of compounding jar (230).

2. The culture device for establishing the Cyr61 target stable transfection cell line according to claim 1, characterized in that: an observation window (111) is installed on the side wall of the box body (110), and a foot seat (112) is connected to the lower wall of the box body (110).

3. The culture device for establishing the Cyr61 target stable transfection cell line according to claim 1, characterized in that: the section of the material guide pipe (210) is arranged in a J shape, and the inner wall of the material guide pipe (210) is a smooth surface.

4. The culture device for establishing the Cyr61 target stable transfection cell line according to claim 1, characterized in that: a lamp holder (231) is installed on the inner side of the upper wall of the mixing tank (230), and an ultraviolet lamp (232) is installed on the lower portion of the lamp holder (231).

5. The culture device for establishing the Cyr61 target stable transfection cell line according to claim 1, characterized in that: the stirring piece (240) comprises a driving motor (241), a supporting shaft (242) and a plurality of stirring blades (243), the driving motor (241) is installed on the upper wall of the mixing tank (230), the supporting shaft (242) is installed at the output end of the driving motor (241), the supporting shaft (242) extends into the inner side of the mixing tank (230), and the stirring blades (243) are installed on the side portions of the supporting shaft (242) respectively.

6. The culture device for establishing the Cyr61 target stable transfection cell line according to claim 5, characterized in that: and scrapers (244) are mounted on the side parts of the stirring blades (243), and the side walls of the scrapers (244) are attached to the inner wall of the mixing tank (230).