CN216473262U

CN216473262U - Medicine test orifice plate suitable for 3D cell ball

Info

Publication number: CN216473262U
Application number: CN202122902751.0U
Authority: CN
Inventors: 余磊; 陈洁琳; 吴万军; 余建辉
Original assignee: Shenzhen Mingao Biotechnology Co ltd
Current assignee: Shenzhen Mingao Biotechnology Co ltd
Priority date: 2021-11-24
Filing date: 2021-11-24
Publication date: 2022-05-10
Anticipated expiration: 2031-11-24

Abstract

The utility model discloses a drug testing pore plate suitable for a 3D cell ball, which comprises a substrate, wherein a plurality of testing holes are arranged on the substrate at intervals, the testing holes are distributed on the substrate in an array manner, the testing holes comprise side walls, side wall holes and bottoms, the substrate comprises one or more than one flow channel, one side of each flow channel is communicated with the adjacent testing hole through the side wall hole, and each flow channel is provided with a driving device. The drive device controls the micro-flow, so that the drug reaction of the tumor is reflected more truly, and the accuracy of drug screening is improved. The utility model can be made into a commercialized pore plate form, can rapidly carry out high-flux test screening on medicines, is suitable for direct measurement and reading of a full-wavelength microplate reader, is beneficial to screening of trace samples, meets the rapid detection requirement of a large number of samples in the market, and has the advantages of high flux, low sample quantity, simple operation and the like.

Description

Medicine test orifice plate suitable for 3D cell ball

Technical Field

The utility model belongs to the field of biomedical materials, and particularly relates to a drug test pore plate suitable for a 3D cell ball.

Background

In vitro cell culture assays are one of the most reliable means of predicting chemotherapeutic drug sensitivity. The potential drug combinations can be validated by drug testing at the cellular level to achieve maximum efficacy and minimal side effects. Since the response of tumor to anticancer drugs is closely related to the microenvironment, careful simulation of tumor microenvironment is the key to obtain accurate drug response information.

Most cells in the human body live in a three-dimensional environment, and they generate physiological functions by obtaining various biochemical and mechanical signals through the close interaction between cells and an extracellular matrix, thereby maintaining their normal physiological activities. Moreover, studies have shown that cell-to-extracellular matrix and cell-to-cell interactions play an important role in regulating cell migration, proliferation and differentiation.

Two-dimensional monolayer cell culture is difficult to truly reflect the living environment of cells in vivo, lacks similar signal transmission, cannot effectively simulate the microenvironment of cells in vivo, and has non-negligible difference with the in vivo. The three-dimensional culture is similar to the in vivo culture in cell types, and simulates the physical communication among cells, so that more real intercellular biological communication is formed.

Therefore, compared with a monolayer cell, the 3D cell can better simulate the microenvironment of cells in vivo, can be more approximate to the growth state of the cells in vivo, is more approximate to the actual situation in various aspects such as gene and function expression, and simultaneously ensures that the cultured cells have more real functions. The result with better in vivo correlation can be obtained by using tumor cell balls formed by 3D cell culture to carry out drug test.

The human body contains fluid transport and molecular diffusion, as well as nanoparticle transport, and cells have internal self-circulation and exchange with the culture environment simultaneously. The existing culture plate or culture chip for experiments is difficult to realize human-like micro-fluidic circulation control, although cells can establish micro-environments in cell nests in semi-closed spaces of the cell nests, the cells need to exchange substances with culture environments, the micro-fluidic needs to be controlled accurately and finely, the micro-environments generated by the cells are damaged when the exchange is too fast, and the support of nutrition is lost when the exchange is too slow.

The required operating procedure of current orifice plate is very loaded down with trivial details, and the middle liquid operation of changing is carried out to and the cell activity in later stage detects, and all steps that relate to the application of sample all accomplish hole by hole, go on in the orifice plate through the manual work, need a large amount of liquid-transfering operations, waste time and energy, can't be applied to the high flux screening of medicine. And the differences between in vivo and in vitro cellular microenvironments have hindered life science research and drug development.

SUMMERY OF THE UTILITY MODEL

Aiming at the defects in the prior art, the utility model aims to provide a drug testing pore plate suitable for 3D cell balls, which is convenient for aggregating cells, can be used for culturing and observing the 3D cell balls and taking images, is suitable for direct measurement and reading of a full-wavelength microplate reader, and realizes high-throughput screening. Meanwhile, the flow channel is arranged on the pore plate, the slow fluid exchange in the cell culture environment is controlled by adopting the micro-fluidic technology under the condition that the culture environment is independent from the culture environment, a more real cell microenvironment is simulated, and a result with better in-vivo correlation can be obtained by utilizing the cell balls to carry out drug testing.

In order to achieve the purpose, the utility model provides the following technical scheme:

the utility model provides a medicine test orifice plate suitable for 3D cell ball, includes a base plate, and the interval has set up a plurality of test holes on the base plate, and the test hole is in be array distribution on the base plate, the test hole contains the lateral wall, lateral wall hole and bottom, and the base plate is provided with one or more runner, and the test hole passes through lateral wall hole and neighbouring the runner intercommunication sets up the drive arrangement of liquid motion in the drive runner on each runner.

Furthermore, the substrate is provided with two or more flow channels, different flow channels are not communicated, the flow channels divide the substrate into two or more independent areas, the testing holes are communicated with the flow channels of the areas through the side wall holes, and each flow channel is provided with a driving device for driving liquid in the flow channel to move. Because the same test orifice plate has two and more than two independent regions, the experimenter often can design the orthogonal experiment based on the experiment purpose, and it can choose the test orifice plate that has a certain number of independent regions for use according to the demand, can carry out the parallel experiment of the different test variables of multiunit simultaneously on same test orifice plate, reduces the loaded down with trivial details operation of hole-by-hole advance kind, also can further reduce the difference that introduces outside the test variable and disturb the test result.

Furthermore, the driving device is a moving rotor arranged in the flow channel and a driving part arranged outside the flow channel, and the driving part outside the flow channel can drive the moving rotor to roll or vibrate, so that the fluid circulation in the driving flow channel can be provided, and the moving rotor is not contacted with the external environment. The size of the drive means is required to be able to roll or vibrate within the flow passage.

Furthermore, the side wall hole is trumpet-shaped, and the aperture of the side wall hole is gradually reduced along the direction from the flow channel to the test hole. The flow velocity is increased when the fluid enters the test hole, and the flow velocity of the microenvironment around the cytosphere is controlled.

Furthermore, the bottom comprises at least one concave surface, and the bottom is of a concave structure, so that cells are convenient to gather, and the cell balling culture and observation are facilitated.

Further, the bottom is transparent for cell observation and image capture.

Further the bottom is a low adhesion or non-adhesion material which may be a perfluorinated polymer, an olefin or similar polymer or a mixture thereof. Avoid the adherence of cells, and is beneficial to the balling culture of the cells. Furthermore, the side wall is opaque, and the side edge of each hole is designed to be black, so that mutual interference between adjacent holes during testing is avoided.

Furthermore, each flow channel on the substrate is provided with a corresponding sample inlet hole, so that the driving device is convenient to take and place for repeated use, and the same concentration to-be-detected medicine is injected into the region communicated with the same flow channel through the sample inlet holes. A recess may be provided in the flow passage for placement of the drive means. Any test hole communicated with the channel can be used as a sample inlet, and a sample inlet hole is not required to be additionally arranged.

Further, the rotor is a magnet with a definite NS pole, and may be a magnet with various shapes, preferably a spherical magnet, and the driving member is a device capable of generating a variable magnetic field externally.

Compared with the prior art, the utility model has the beneficial effects that: the application of the utility model solves the problems of complicated operation and poor correlation of the traditional in-vitro drug screening method, and the cultured 3D tumor cells can be used for high-flux antitumor drug evaluation and candidate drug screening test. The problems that the micro-environment in vivo is not considered in the traditional culture method are solved, the micro-flow is controlled by the magnetons, the drug reaction of the tumor is reflected more truly, and the accuracy of drug screening is improved.

The utility model can be made into a commercialized pore plate form, can rapidly carry out high-flux test screening on medicines, is suitable for direct measurement and reading of a full-wavelength microplate reader, is beneficial to screening of trace samples, meets the rapid detection requirement of a large number of samples in the market, and has the advantages of high flux, low sample quantity, simple operation and the like. Has great application prospect in the fields of tumor research, anti-tumor drug development and the like.

Drawings

FIG. 1 is a schematic partial cross-sectional view of an embodiment of the present invention;

FIG. 2 is an enlarged schematic view of a portion A of FIG. 1;

FIG. 3 is a schematic top view of an embodiment of the present invention;

FIG. 4 is a schematic top view of the structure of FIG. 1;

FIG. 5 is a cross-sectional view taken along line F-F of FIG. 4, the sidewalls being opaque and the base being mounted on the substrate;

FIG. 6 is a schematic cross-sectional view taken along line F-F of FIG. 4, with the bottom portion coated with a low adhesion material and the bottom portion exposed to the external environment;

fig. 7 is a schematic cross-sectional view taken along line F-F of fig. 4, showing opaque sidewalls and a bottom exposed to the external environment.

In the figure, 1, a substrate; 2. a test well; 21. a side wall; 22. a sidewall hole; 23. a bottom; 3. a flow channel; 4. a driving device (mover); 5. and (4) sampling holes.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.

Referring to fig. 1 to 7, an embodiment of the present invention is:

in the first embodiment, a drug test well plate suitable for 3D cell balls is provided, in which 12 × 8 test wells 2 are arranged in an array on a substrate 1, and 96 test wells 2 are provided in total. The base plate 1 is provided with a flow channel 3, the test hole 2 is communicated with the flow channel 3 through a side wall hole 22, and the spherical magneton 4 is arranged in the flow channel 3.

The 3D cell ball is placed in the test hole firstly, then the medicine to be tested is prepared into liquid, and the liquid is injected into the flow channel 3 through the sample injection hole 5, so that the complicated operation of sample injection from hole to hole is reduced. Finally, a magnetic field changing device arranged outside is used for generating a changing magnetic field to continuously drive the spherical magneton 4 to roll, rotate or linearly reciprocate so as to form a micro-circulation flow field. During the test, the 3D cell balls are in the micro-circulation flow field, so that the cells not only have internal self-circulation, but also exchange with the culture environment at the same time.

The second embodiment provides a drug test well plate suitable for 3D cell balls, in which 24 × 16 test wells 2 are arranged in an array on a substrate 1, and a total of 384 test wells 2 are provided. The substrate 1 is provided with a flow channel 3, the substrate 1 is divided into 8 independent test areas through the flow channel 3, the test hole 2 in each area is communicated with the flow channel 3 in the area through a side wall hole 22 and is not communicated with other flow channels 3, and a spherical magneton 4 is arranged in each flow channel 3. According to the experimental requirements, the test orifice plate can be designed into a certain number of test orifices 2 and divided into a certain number of independent areas by using a certain number of flow channels 3, and the specific number is not limited by the embodiment.

Firstly, 3D cell balls are placed in the testing holes 2, and then according to the experimental design of drug testing, drugs to be tested are configured into liquids with different concentrations and respectively injected into different flow channels 3. The magnetic field changing device arranged outside generates a changing magnetic field to continuously drive the spherical magneton 4 to roll, rotate or linearly reciprocate to form a micro-circulation flow field. During the test, the 3D cell balls are in the micro-circulation flow field, so that the cells not only have internal self-circulation, but also exchange with the culture environment at the same time. Because the test orifice plate contains a plurality of independent test areas, a plurality of groups of parallel experiments can be simultaneously carried out on the same test orifice plate.

Example three provides a drug test well plate suitable for 3D cell balls, in which 24 × 16 test wells 2 are arranged in an array on a substrate 1, and a total of 384 test wells 2 are provided. The substrate 1 is provided with a flow channel 3, the test hole 2 is communicated with the flow channel 3 through a side wall 21 hole, and the bottom 23 of the test hole 2 is of a concave structure, so that cells are convenient to gather, and the cell balling culture and observation are facilitated. A bar-shaped magnet 4 is provided in the flow path 3.

The bar-shaped magnetons 4 are continuously driven to vibrate and reciprocate by the variable magnetic field generated by the magnetic field change device arranged outside to form a micro-circulation flow field. During testing, the 3D cell spheres were in a micro-circulation flow field, and the cells had internal self-circulation and were simultaneously exchanged with the culture environment.

Example four provides a drug-testing well plate suitable for 3D cell balls, on a substrate 1, 24 × 16 test wells 2 are arranged in an array, totaling 384 test wells 2. The base plate 1 is provided with a flow channel 3, and the test hole 2 is communicated with the flow channel 3 through a side wall hole 21. The bottom 23 of the test well 2 is transparent and the bottom 23 is exposed to the external environment for cell observation and image capture. A spherical magnet 4 is provided in the flow channel 3.

Example five provides a drug-testing well plate suitable for 3D cell balls, on which 24 × 16 test wells 2 are arranged in an array on a substrate 1, for a total of 384 test wells 2. The base plate 1 is provided with a flow channel 3, and the test hole 2 is communicated with the flow channel 3 through a side wall 21 hole. The lateral wall 21 of test hole 2 designs for the black, avoids mutual interference between the adjacent test hole 2 when testing, and the bottom 23 of test hole 2 inlays in base plate 1, utilizes base plate 1 protection test hole 2's bottom 23. A spherical magnet 4 is provided in the flow channel 3.

In the sixth embodiment, a drug test well plate suitable for 3D cell balls is provided, in which 24 × 16 test wells 2 are arranged in an array on a substrate 1, and a total of 384 test wells 2 are provided. The base plate 1 is provided with a flow channel 3, and the test hole 2 communicates with the flow channel 3 through a side wall hole 22. The bottom 23 of the test hole 2 is coated with a low adhesion material, the low adhesion coating is perfluorinated polymer, and the spherical magnetons 4 are arranged in the flow channels 3. Other embodiments of the low adhesion coating may be an olefin or similar polymer or mixtures thereof.

The above description is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the utility model may occur to those skilled in the art without departing from the principle of the utility model, and are considered to be within the scope of the utility model.

Claims

1. A drug test well plate suitable for 3D cell balls, characterized in that: the testing device comprises a substrate, a plurality of testing holes have been set up at the interval on the base plate, the testing hole is in be array distribution on the base plate, the testing hole contains lateral wall, lateral wall hole and bottom, the base plate is provided with one or more runner, the testing hole passes through lateral wall hole and neighbouring the runner intercommunication, each set up the drive on the runner the drive arrangement of liquid motion in the runner.

2. A drug testing well plate suitable for use in 3D cell pellets according to claim 1, wherein: the base plate is provided with two or more than two the runner, it is different not communicate between the runner, the runner will the base plate is separated into two or more than independent regions, the test hole passes through side wall hole and place regional the runner intercommunication, each set up the drive on the runner liquid motion's in the runner drive arrangement.

3. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: the driving device is a moving rotor arranged in the flow channel and a driving part arranged outside the flow channel, and the moving rotor can be driven to roll or vibrate by the driving part outside the flow channel.

4. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: the side wall hole is trumpet-shaped, and the aperture of the side wall hole is gradually reduced along the direction from the flow channel to the test hole.

5. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: the base comprises at least one concave surface.

6. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: the bottom is transparent.

7. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: the bottom is of low or no adhesion material.

8. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: the side walls are opaque.

9. A drug test well plate suitable for use in 3D cell spheres according to claim 1 or 2, wherein: each flow channel on the substrate is provided with a corresponding sample inlet hole.

10. A drug testing well plate suitable for use with 3D cell spheres, according to claim 3, wherein: the moving rotor is a spherical magnet with definite NS pole, and the driving part is a device capable of generating variable magnetic field externally.