A cervical cancer stem cell culture device

Abstract

This invention belongs to the technical field of stem cell culture equipment, specifically relating to a cervical cancer stem cell culture device, comprising a cover, a dish, and a bottom cavity. The cover is detachably fitted onto the top of the dish, and the bottom of the dish has a bottom cavity. The dish and the bottom cavity are connected by a first pipe, which is equipped with a valve. The cover has a stretchable and deformable receiving surface, and a second pipe is connected to the cover above the receiving surface. One end of the second pipe is connected to fresh sterile culture medium. The receiving surface has an outlet, which changes between closed and open states as the shape of the receiving surface changes. This invention provides a cervical cancer stem cell culture device, which is a closed cervical cancer stem cell culture device that avoids environmental pollution while meeting the culture requirements of cervical cancer stem cells.

Description

Technical Field

[0001] This invention belongs to the field of stem cell culture equipment technology, specifically relating to a cervical cancer stem cell culture device. Background Technology

[0002] Cancer stem cells refer to cancer cells that possess stem cell-like properties, meaning they have the ability to self-replicate and differentiate into multiple cells. The characteristics of tumor cell growth, metastasis, and recurrence are very similar to the basic properties of stem cells. Therefore, some scholars have proposed the theory of cancer stem cells. This theory provides a new direction and perspective for our re-understanding of the origin and nature of tumors, as well as clinical cancer treatment. The difference between cancer stem cells and ordinary tumor cells is that cancer stem cells are like "seeds of tumors." Tumors can grow indefinitely, but not every tumor cell can grow indefinitely; rather, it is the cancer stem cells within them that grow indefinitely and can travel through the bloodstream to any healthy site to take root and sprout into new tumors. Ordinary tumor cells cannot do this. This is the key difference. Therefore, research on cancer stem cells is of great significance for cancer treatment.

[0003] Studies have shown that millions of ordinary tumor cells are needed to induce tumor formation in mice, while only 100 tumor stem cells are required. Furthermore, since tumor stem cells can grow indefinitely, the process of culturing and researching them requires even greater attention to prevent contamination compared to ordinary tumor cells.

[0004] Cervical cancer stem cells are a type of tumor stem cell, such as the cervical cancer stem cell spheroids isolated by Zhang Ying et al. (Zhang Ying, Chang Yanan, Liu Huan, et al. Screening and identification of stem cell spheroids in cervical cancer cell lines [J]. Cancer Progress, 2014(6):6.DOI:10.11877 / j.issn.1672-1535.2014.12.06.10.). Zhang Ying et al. also disclosed the specific culture method of cervical cancer stem cell spheroids, including: trypsin digestion to collect Ishikawa cells grown to 90% density, washing once with PBS and resuspending with SCM, counting, and seeding in Corning ultra-low adhesion six-well plates with a cell concentration of 1000 / ml. After culturing for 7-9 days, tumor cell spheres were collected in 15ml centrifuge tubes and allowed to settle naturally for 10 minutes. The supernatant was then aspirated to remove cell debris. After washing once with PBS, Accutase was added and the tubes were placed in an incubator for digestion for 5-10 minutes, during which time the cells were gently pipetted with a 1ml pipette tip intermittently. After digestion, SCM was added and the cells were passaged in an ultra-low adhesion plate at a rate of 1000 / ml.

[0005] The culture device used in the above-mentioned cervical cancer stem cell culture process is a multi-well plate, which includes multiple culture wells and the culture wells are exposed to the outside. When the cervical cancer stem cell culture process involves multiple "aspiration and supernatant removal" processes, the exposed environment poses a risk of leakage of cervical cancer stem cells and environmental pollution. Therefore, it is necessary to provide a closed cervical cancer stem cell culture device. Summary of the Invention

[0006] To address the aforementioned technical problem that "the culture wells of multi-well plates used for cervical cancer stem cells are exposed to the outside, posing a risk of leakage and environmental pollution of cervical cancer stem cells," this invention provides a cervical cancer stem cell culture device, which is a closed cervical cancer stem cell culture apparatus that can both avoid environmental pollution and meet the culture requirements of cervical cancer stem cells.

[0007] The purpose of this invention is to provide a cervical cancer stem cell culture device, comprising a cover, a dish, and a bottom cavity, wherein the cover is detachably fitted onto the top of the dish, and the bottom cavity is provided at the bottom of the dish;

[0008] The dish body is connected to the bottom cavity via a first pipe, and a valve is provided on the first pipe;

[0009] The cover body has a stretchable and deformable receiving surface. A second pipe is connected to the cover body above the receiving surface. One end of the second pipe is connected to fresh sterile culture medium, and the other end is connected to the cover body. Fresh sterile culture medium can flow into the receiving surface through the second pipe. The receiving surface has a liquid outlet. When the weight of the culture medium on the receiving surface is insufficient to cause the receiving surface to stretch and deform downward, the liquid outlet is closed. When the weight of the culture medium on the receiving surface is sufficient to cause the receiving surface to stretch and deform downward, the liquid outlet is open.

[0010] Preferably, in the above-mentioned cervical cancer stem cell culture device, the thickness of the receiving surface is set to less than 1 mm, and the material of the receiving surface is an elastic waterproof membrane.

[0011] Preferably, in the above-mentioned cervical cancer stem cell culture device, the highest point of the first pipe is located at 1 / 2 to 2 / 3 of the height of the dish body 2.

[0012] Preferably, in the above-mentioned cervical cancer stem cell culture device, the receiving surface includes a first telescopic surface and a second telescopic surface spliced ​​together, the position between the opposite edges of the first telescopic surface and the second telescopic surface is the liquid outlet, and the other edges of the first telescopic surface and the second telescopic surface are connected to the inner wall of the cover body adjacent to each other.

[0013] Preferably, in the above-mentioned cervical cancer stem cell culture device, the edges of the first and second telescopic surfaces are provided with adhesive strips, the adhesive strips of the first and second telescopic surfaces attract each other, and the position between the two adhesive strips is the liquid outlet.

[0014] Preferably, in the above-mentioned cervical cancer stem cell culture device, the adhesive strip is provided with an adsorption block, and the bottom of the dish is provided with an adsorbent that attracts the adsorption block.

[0015] Preferably, in the above-mentioned cervical cancer stem cell culture device, the bottom cavity is provided with a deformable barrier surface, which divides the bottom cavity into a culture medium storage cavity and a waste liquid storage cavity. The first pipe is connected and communicates with the waste liquid storage cavity, and the culture medium storage cavity is connected and communicates with the second pipe. The cavity walls of both the culture medium storage cavity and the waste liquid storage cavity are provided with material flow ports, and both material flow ports are provided with sealing caps. The second pipe is provided with a liquid extraction device.

[0016] Preferably, in the above-mentioned cervical cancer stem cell culture device, the sealing cover is provided with a flow hole, the flow hole is sealed with an air-permeable and water-permeable component, and the position of the flow hole outside the air-permeable and water-permeable component is sealed with a bacterial filter component.

[0017] Preferably, the above-mentioned cervical cancer stem cell culture device further includes a height adjustment device, which includes a sealing component. The side wall of the dish body has an installation opening, and the sealing component is detachably fitted into the installation opening. The sealing component has a through hole, one end of the first pipe is a curved pipe, and a sealing component is sleeved on the outside of the curved pipe. After the sealing component is embedded in the through hole, the curved pipe passes through the through hole. The sealing component is elastic, and the position of the curved pipe inside the sealing component is adjustable.

[0018] Preferably, in the above-mentioned cervical cancer stem cell culture device, the height adjustment device further includes a clamping component, which is installed on the sealing component, and the curved tube is clamped by the clamping component.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The dish of this invention serves as the culture site for cervical cancer stem cells, with both the culture medium and the cervical cancer stem cells placed inside. The bottom cavity is for collecting waste supernatant. The cervical cancer stem cell culture equipment is placed inside an incubator, which provides the necessary temperature, humidity, and other conditions for cell growth. The incubator utilizes existing technology, such as replacing existing culture flasks. A lid is fitted over the dish, and the dish is connected to the bottom cavity via a first conduit, while the lid is connected to the bottom cavity via a second conduit, forming a closed space for culture and medium replacement, preventing leakage of cervical cancer stem cells.

[0021] After opening the valve on the first pipe, gravity allows the supernatant in the dish to flow into the bottom cavity through the first pipe. The space below the receiving surface is used to place the cervical cancer stem cells to be cultured and the culture medium. When the cervical cancer stem cells have been cultured to a certain extent and the medium needs to be changed, the valve on the first pipe is opened, and the supernatant to be discarded flows from the first pipe to the bottom cavity. Meanwhile, fresh sterile culture medium is added to the top of the receiving surface through the second pipe. Since the outlet of the receiving surface is closed at this time, and the space between the receiving surface and the dish is sealed, as fresh sterile culture medium is continuously added to the top of the receiving surface, the receiving surface gradually stretches and deforms downwards. This causes the receiving surface to exert a compressive force on the material in the space between the receiving surface and the dish. This compressive force accelerates the flow of the waste supernatant from the first pipe to the bottom cavity. When the volume of culture medium on the receiving surface is insufficient to open the outlet, the fresh sterile culture medium remains temporarily above the receiving surface and does not flow into the bottom cavity with the waste supernatant, thus avoiding waste of fresh sterile culture medium. When the volume of culture medium on the receiving surface is sufficient to open the outlet, fresh sterile culture medium flows into the dish. At this point, the receiving surface, under the action of elasticity, restores the original shape of the outlet and closes again.

[0022] Based on the above principles, this invention achieves a seamless process of discarding the supernatant from the dish and adding fresh sterile culture medium. Both the "discarding of supernatant" and the "replacing with fresh sterile culture medium" are carried out in a closed environment, thus avoiding contamination of the external environment by tumor stem cells. Attached Figure Description

[0023] Figure 1 This is a three-dimensional structural diagram of the cervical cancer stem cell culture device of the present invention.

[0024] Figure 2 The working principle of the cervical cancer stem cell culture device of the present invention Figure 1 (The dashed lines represent components inside the lid, dish, or bottom cavity).

[0025] Figure 3 The working principle of the cervical cancer stem cell culture device of the present invention Figure 2 (The dashed lines represent components inside the lid, dish, or bottom cavity).

[0026] Figure 4 This is a top view of the receiving surface of the present invention.

[0027] Figure 5 This is a schematic diagram showing the connection between the vessel body, the bottom cavity, and the first pipe of the present invention.

[0028] Figure 6 This is a schematic diagram of the height adjustment device of the present invention. Detailed Implementation

[0029] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0030] Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0031] In the description of this invention, it should be understood that the terms "center," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," and "circumferential" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0032] The terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature; in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0033] Example 1

[0034] This invention provides a cervical cancer stem cell culture device, see [link to device]. Figure 1The device includes a lid 1, a dish 2, and a bottom cavity 3. The top of the dish 2 is open, and the bottom is closed. The lid 1 can be placed on top of the dish 2, sealing the top opening. The lid 1 can also be removed from the dish 2. Preferably, the part of the dish 2 used to connect to the lid 1 has a sealing ring, so that after the lid 1 is placed on top of the dish 2, the connection between the two can be sealed to prevent leakage and leakage of cervical cancer stem cells. The bottom of the dish 2 has a bottom cavity 3, and the dish 2 is placed or fixedly connected to the upper surface of the bottom cavity 3. The dish 2 is the place for culturing cervical cancer stem cells, and both the culture medium and cells are placed inside the dish 2. The bottom cavity 3 is the place for collecting waste supernatant. The cervical cancer stem cell culture equipment is placed in an incubator, which provides the temperature, humidity, and other conditions required for cell growth. The incubator adopts existing technology, such as an existing incubator capable of holding culture flasks / multi-well plates.

[0035] Continue to refer to Figure 1 The dish body 2 and the bottom cavity 3 are connected by a first pipe 4, which is equipped with a valve 41. Preferably, the first pipe 4 is located outside the dish body 2 and the bottom cavity 3. The highest point of the first pipe 4 is lower than the liquid level in the dish body 2, especially lower than the highest liquid level of the supernatant. When the valve 41 is opened, the supernatant in the dish body 2 can enter the bottom cavity 3 through the first pipe 4 due to gravity. The amount of culture medium added to the dish body 2 needs to be manually controlled. The liquid level of the culture medium is higher than the opening where the first pipe 4 connects to the dish body 2, and the opening where the first pipe 4 connects to the dish body 2 is the highest point of the first pipe 4. Of course, in order to easily control the liquid level of the culture medium in the dish body 2, the highest point of the first pipe 4 cannot be too high. Preferably, the highest point of the first pipe 4 is located at 1 / 2 to 2 / 3 of the height of the dish body 2.

[0036] Reference Figure 2 The cover 1 has a stretchable and deformable receiving surface 11. A distance is left between the receiving surface 11 and the inner top wall of the cover 1. After the cover 1 is connected to the dish 2, the space below the receiving surface 11 is used to place the cervical cancer stem cells to be cultured and the culture medium. Cervical cancer stem cells have their own specific culture medium components, such as trypsin, DMEM / F12 medium, fetal bovine serum, recombinant human epidermal growth factor, and fibroblast growth factor used by Zhang Ying et al. in the background art of this invention. A second pipe 5 is connected to the cover 1 above the receiving surface 11. One end of the second pipe 5 is connected to fresh sterile culture medium, and the other end is connected to the inside of the cover 1. Thus, the fresh sterile culture medium enters the cover 1 through the second pipe 5 and falls onto the receiving surface 11. (Refer to...) Figure 2 The receiving surface 11 has a liquid outlet 111. When the weight of the culture medium on the receiving surface 11 is insufficient to cause the receiving surface 11 to undergo downward stretching deformation, the liquid outlet 111 is closed, and at this time, there is no leakage at the liquid outlet 111. (Refer to...) Figure 3 When there is enough culture medium on the receiving surface 11, and the weight of the culture medium is sufficient to cause the receiving surface 11 to stretch and deform downward, the outlet 111 is open. At this time, the outlet 111 leaks liquid, and the culture medium flows to the bottom of the receiving surface 11.

[0037] It should be noted that preliminary experiments should be conducted beforehand to determine the relationship between the volume of the culture medium and the timing of the downward stretching deformation of the receiving surface 11, as well as the material and thickness of the receiving surface 11. For example, when the culture medium above the receiving surface 11 reaches 100 μL, 1 mL, or 10 mL, the receiving surface 11 will undergo downward stretching deformation, opening the outlet 111. When the culture medium above the receiving surface 11 is below a certain volume, the outlet 111 will be closed. Since the volume of culture medium required for cervical cancer stem cells is usually not large, the thickness of the receiving surface 11 is set to less than 1 mm, and the material of the receiving surface 11 is chosen to be an elastic latex film or other elastic waterproof film.

[0038] The working process of this invention includes the following steps:

[0039] First, the configuration device: according to Figure 1 The structure shown includes a cover 1, a dish 2, a bottom cavity 3, a first pipe 4, and a second pipe 5. The dish 2 is positioned on the bottom cavity 3. The first pipe 4 connects the dish 2 to the bottom cavity 3, and the second pipe 5 connects to the cover 1. Valve 41 is closed. To improve the sealing effect, the inlet of the second pipe 5, used for connecting fresh sterile culture medium, is equipped with a filter plug, such as sterile cotton.

[0040] Second, inoculate cervical cancer stem cells and culture medium into dish 2, ensuring that the height of the adherent cervical cancer stem cells is lower than the lowest point of the opening of the first conduit 4 connecting to dish 2. Cover dish 2 with cap 1, thus creating a relatively closed environment for the cervical cancer stem cells and culture medium; then place the assembled equipment into an incubator for culturing.

[0041] Third, when the cervical cancer stem cells have been cultured to a certain extent and the medium needs to be changed, open valve 41 of the first pipe 4. The supernatant to be discarded will flow from the first pipe 4 to the bottom cavity 3. Additionally, remove the filter plug from the second pipe 5 and add fresh sterile culture medium above the receiving surface 11 via the second pipe 5. Since the outlet 111 of the receiving surface 11 is closed at this time, the space between the receiving surface 11 and the dish body 2 is sealed. As fresh sterile culture medium is continuously added above the receiving surface 11, the receiving surface 11 gradually extends downwards. At this time, the receiving surface 11 exerts a squeezing effect on the material in the space between the receiving surface 11 and the dish body 2. This squeezing effect accelerates the flow of the waste supernatant from the first pipe 4 to the bottom cavity 3, similar to the dispensing principle of a syringe.

[0042] Furthermore, when the volume of culture medium on the receiving surface 11 is insufficient to open the outlet 111, the fresh sterile culture medium remains temporarily above the receiving surface 11 and will not flow into the bottom cavity 3 with the waste supernatant, thus avoiding waste of fresh sterile culture medium. Therefore, the fresh sterile culture medium flows into the dish 2 later than the waste supernatant flows into the bottom cavity 3. When the volume of culture medium on the receiving surface 11 is sufficient to open the outlet 111, the waste supernatant has almost completely flowed into the bottom cavity 3. At this time, the outlet 111 opens, referring to... Figure 3 Fresh, sterile culture medium entered the dish 2, and then, under the action of elasticity, the receiving surface restored the original shape of the outlet 111, closing again, and returned to its original state. Figure 2 The location.

[0043] Based on the above principles, this invention achieves a seamless process of discarding the supernatant from the dish 2 and adding fresh sterile culture medium. Both the "discarding of supernatant" and the "replacing with fresh sterile culture medium" are carried out in a closed environment, thus avoiding contamination of the external environment by tumor stem cells.

[0044] For example, refer to Figure 2-4 The receiving surface 11 includes a first telescopic surface and a second telescopic surface joined together. The area between the opposite edges of the first and second telescopic surfaces forms the outlet 111. The other edges of the first and second telescopic surfaces are fixedly connected to the inner wall of their respective adjacent caps 1. The first and second telescopic surfaces are made of the same material as the receiving surface 11. Each of the opposite edges of the first and second telescopic surfaces is provided with an adhesive strip 112. The adhesive strips 112 of the first and second telescopic surfaces attract each other, and the area between the two adhesive strips 112 forms the outlet 111.

[0045] For example, the bonding strip 112 is a structure such as a magnetic strip or an electrostatic strip that can attract each other.

[0046] For example, the adhesive strip 112 is provided with an adsorption block, and the bottom of the dish body 2 is provided with an adsorbent 21 that attracts the adsorption block. As the culture medium on the receiving surface 11 becomes larger, the receiving surface 11 extends downward, that is, towards the bottom of the dish body 2. That is, when the first and second extension surfaces extend, the distance between the adsorption block on the adhesive strip 112 and the adsorbent 21 gradually approaches. When the adsorption block reaches a certain position, the adsorbent 21 has sufficient attraction to the adsorption block. In addition, the gravity of the culture medium on the receiving surface 11 results in a large force on the adhesive strip 112 at the outlet 111, which facilitates the smooth opening of the outlet 111.

[0047] For example, the adsorption block is made of iron or other materials that can be attracted to a magnet, and the adsorption body 21 is a magnet. According to the principle of magnetic attraction, the closer to the magnet, the stronger the magnetic attraction.

[0048] Continue to refer to Figure 2-3 To facilitate the addition of fresh sterile culture medium and further enhance the "sealing effect" of the device, a deformable barrier surface 31 is provided in the bottom cavity 3. For example, the barrier surface 31 can be made of a waterproof latex or rubber material. The barrier surface 31 divides the bottom cavity 3 into a culture medium storage cavity 32 and a waste liquid storage cavity 33. The culture medium storage cavity 32 is located above or below the waste liquid storage cavity 33. The first pipe 4 is connected to and communicates with the waste liquid storage cavity 33, and the connection seam between the two is sealed with a sealing gasket to prevent leakage. The culture medium storage cavity 32 is connected to and communicates with the second pipe 5, and the connection seam between the two is sealed with a sealing strip to prevent leakage. Fresh sterile culture medium is stored in the culture medium storage cavity 32. With the culture medium storage cavity 32, there is no need to install a filter plug.

[0049] Both the culture medium storage chamber 32 and the waste liquid storage chamber 33 have material flow ports on their walls, and each material flow port is equipped with a sealing cap 34. When the sealing cap 34 is closed, the material flow ports are sealed, and both the culture medium storage chamber 32 and the waste liquid storage chamber 33 are in a sealed state. Therefore, when the waste supernatant enters the waste liquid storage chamber 33 through the first pipe 4, the pressure inside the waste liquid storage chamber 33 increases, pressing the barrier surface 31 towards the culture medium storage chamber 32. Thus, the barrier surface 31 bulges towards the culture medium storage chamber 32. (See [reference]). Figure 3 The culture medium in the culture medium storage cavity 32 is squeezed and forced into the second pipe 5, eventually flowing into the cover 1. To make this flow smoother, a pumping device 51, such as a pump, is provided on the second pipe 5. After the pumping device 51 is turned on, the culture medium in the culture medium storage cavity 32 flows into the cover 1 under the combined effect of gravity and the waste supernatant. This makes it easy for either the waste supernatant to flow into the waste liquid storage cavity 33 or for fresh sterile culture medium to flow into the cover 1. Furthermore, due to the gravity of the waste supernatant, the power consumption of the pumping device 51 can be reduced, saving energy.

[0050] For example, the edge of the barrier surface 31 is fixedly connected to the inner wall edge of the bottom cavity 3.

[0051] When it is necessary to adjust the pressure inside the bottom cavity 3, a flow hole is provided on the sealing cover 34. The flow hole is closed with a breathable but waterproof component, such as a breathable but waterproof membrane, so that the pressure inside the bottom cavity 3 is balanced by gas flow. At this time, the position of the flow hole outside the bottom cavity 3 is sealed with a filter component to prevent contamination. The filter component is filter cotton.

[0052] For example, a vent hole can be made on the top or side wall of the cover 1 above the receiving surface 11. A cell group septum is provided at the vent hole. The cell group septum is a filter membrane or other form of membrane. The purpose is to allow air to pass through, but not to allow cervical cancer stem cells to pass through. This can prevent cell leakage and also allow the gas environment inside the incubator to communicate with the environment inside the dish 2 when the outlet 111 is opened, so as to provide a suitable gas environment for the growth of cervical cancer stem cells.

[0053] To facilitate the flow of waste supernatant from dish 2 to the first conduit 4 while preventing cervical cancer stem cells from dish 2 from flowing into the first conduit 4, this invention includes a height adjustment device. The height adjustment device is used to adjust the height of the opening of the first conduit 4 within dish 2.

[0054] See Figure 5-6 The height adjustment device includes a sealing component 6. An installation opening is provided on the side wall of the dish body 2, and the sealing component 6 is detachably fitted into the installation opening. For example, the sealing component 6 can be an elastic block that can be embedded in the installation opening and has both waterproof and sealing functions, preventing leakage from the installation opening. The sealing component 6 has a through hole, and one end of the first pipe 4 is a curved tube. A sealing component 42 is fitted onto the outside of the curved tube. After the sealing component 42 is embedded in the through hole, the curved tube passes through the through hole. The sealing component 42 is elastic; for example, it can be an elastic membrane or an elastic block made of latex or rubber. The position of the curved tube within the sealing component 42 is adjustable. For example, when the curved tube moves inwards towards the inside of the dish body 2, that is, from... Figure 5 If the bend moves from right to left, then under the influence of "curvature," the position of the bend's opening inside the dish 2 decreases. When the bend moves outside the dish 2, that is, from... Figure 5 If the tube is moved from left to right, the position of the curved tube opening within dish 2 will rise due to the influence of the "curvature". Therefore, when the volume of newly added fresh sterile culture medium in dish 2 changes and its height in dish 2 changes, this change can be accommodated by adjusting the height of the curved tube opening.

[0055] For example, the bending angle of the bent tube is 20-40 degrees.

[0056] The height adjustment device also includes a clamping component 7, which is mounted on the sealing component 6. The bent tube is clamped and limited by the clamping component 7.

[0057] For example, the clamping component 7 includes a first clamping part and a second clamping part, both of which are slidably connected to the sealing component 6. The curved tube also passes through the space between the first clamping part and the second clamping part. When the first clamping part and the second clamping part move toward each other and connect, the curved tube can be clamped.

[0058] For example, the sealing component 6 has a groove, and both the first clamping part and the second clamping part are provided with sliders that are slidably connected to the groove. The opposing surfaces of the first clamping part and the second clamping part are provided with connecting members. When the bent tube passes through the space between the first clamping part and the second clamping part, the connecting members can also connect, thereby tightly clamping the first clamping part and the second clamping part outside the bent tube. Preferably, the groove is vertically arranged to prevent the first clamping part and the second clamping part from rotating erratically, thus preventing the bent tube from rotating erratically.

[0059] For example, the connecting component is a detachable connection such as a snap-fit ​​or tenon joint.

[0060] It should be noted that the connection relationships of components not specifically mentioned in this invention are all assumed to be based on existing technology. Since they do not involve the inventive point and are commonly used in existing technology, the structural connection relationships are not described in detail.

[0061] It should be noted that when numerical ranges are involved in this invention, it should be understood that both endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as in the embodiments, preferred embodiments are described here to avoid redundancy. Although preferred embodiments of the invention have been described, those skilled in the art, once they understand the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of this invention.

[0062] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A cervical cancer stem cell culture device, characterized in that, It includes a cover (1), a dish (2) and a bottom cavity (3). The cover (1) is detachably fitted onto the top of the dish (2), and the bottom cavity (3) is provided at the bottom of the dish (2). The dish body (2) and the bottom cavity (3) are connected by a first pipe (4), and a valve (41) is provided on the first pipe (4). The cover (1) has a stretchable and deformable receiving surface (11) inside. A second pipe (5) is connected to the cover (1) above the receiving surface (11). One end of the second pipe (5) is connected to a fresh sterile culture medium, and the other end of the second pipe is connected to the cover (1). The fresh sterile culture medium can flow into the receiving surface (11) through the second pipe (5). The receiving surface (11) has an outlet (111). When the weight of the culture medium on the receiving surface (11) is insufficient to cause the receiving surface (11) to stretch downward, the outlet (111) is closed. When the weight of the culture medium on the receiving surface (11) is sufficient to cause the receiving surface (11) to stretch downward, the outlet (111) is open. The receiving surface (11) includes a first telescopic surface and a second telescopic surface spliced ​​together. The position between the opposite edges of the first telescopic surface and the second telescopic surface is the liquid outlet (111). The other edges of the first telescopic surface and the second telescopic surface are connected to the inner wall of the cover (1) adjacent to each other. The edges of the first and second telescopic surfaces are provided with adhesive strips (112). The adhesive strips (112) of the first and second telescopic surfaces attract each other, and the position between the two adhesive strips (112) is the liquid outlet (111).

2. The cervical cancer stem cell culture device according to claim 1, characterized in that, The thickness of the receiving surface (11) is set to less than 1 mm, and the material of the receiving surface (11) is an elastic waterproof membrane.

3. The cervical cancer stem cell culture device according to claim 1, characterized in that, The highest point of the first pipe (4) is located at 1 / 2 to 2 / 3 of the height of the dish (2).

4. The cervical cancer stem cell culture device according to claim 1, characterized in that, The adhesive strip (112) is provided with an adsorption block, and the bottom of the dish body (2) is provided with an adsorbent (21) that attracts the adsorption block.

5. The cervical cancer stem cell culture device according to claim 1, characterized in that, The bottom cavity (3) is provided with a deformable barrier surface (31), which divides the bottom cavity (3) into a culture medium storage cavity (32) and a waste liquid storage cavity (33). The first pipe (4) is connected to the waste liquid storage cavity (33), and the culture medium storage cavity (32) is connected to the second pipe (5). The walls of the culture medium storage cavity (32) and the waste liquid storage cavity (33) are provided with material flow ports, and sealing caps (34) are provided at both material flow ports. A liquid extraction device (51) is provided on the second pipe (5).

6. The cervical cancer stem cell culture device according to claim 5, characterized in that, The sealing cap (34) is provided with a flow hole, which is sealed with a breathable but waterproof component, and the position of the flow hole outside the breathable but waterproof component is sealed with a bacteria-filtering component.

7. The cervical cancer stem cell culture device according to claim 1, characterized in that, It also includes a height adjustment device, which includes a sealing component (6). The side wall of the dish body (2) has an installation opening. The sealing component (6) is detachably fitted into the installation opening. The sealing component (6) has a through hole. One end of the first pipe (4) is a curved pipe. A sealing component (42) is fitted on the outside of the curved pipe. After the sealing component (42) is embedded in the through hole, the curved pipe passes through the through hole. The sealing component (42) is elastic, and the position of the curved pipe in the sealing component (42) is adjustable.

8. The cervical cancer stem cell culture device according to claim 7, characterized in that, The height adjustment device also includes a clamping component (7), which is mounted on the sealing component (6), and the curved tube is clamped by the clamping component (7).

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