In-vitro osteosarcoma culture chip

Abstract

The utility model provides an osteosarcoma in-vitro culture chip which comprises a carrier plate, an outlet hole and at least two inlet holes are formed in the two ends of the carrier plate respectively, the inlet holes are higher than the outlet hole, and a mixing channel area, a concentration gradient forming area, a tissue culture area and a liquid storage buffer area which are communicated and are gradually decreased in height are sequentially arranged between the inlet holes and the outlet holes. The concentration mixing channel area comprises a mixing channel, inflow ends and outflow ends which are communicated, the inflow ends and the outflow ends are arranged on the two sides of the mixing channel respectively, the inflow ends are arranged between the adjacent outflow ends in the length direction of the mixing channel, and the number of the outflow ends of the concentration mixing channel area is one more than that of the inflow ends. The multiple concentration gradient forming areas are arranged side by side, all the inlet holes are communicated with all the inflow ends through the flow channels, all the outflow ends are communicated with all the concentration gradient forming areas through the flow channels, and the problem that the medicine concentration screening efficiency is low during osteosarcoma in-vitro culture is solved.

Description

Technical Field

[0001] This utility model relates to the field of osteosarcoma in vitro culture, and in particular to an osteosarcoma in vitro culture chip. Background Technology

[0002] In recent years, organ-on-a-chip technology, as an emerging in vitro model, has gradually become an important tool for studying the tumor microenvironment, drug screening, and biological response mechanisms in tissue engineering and cancer biology research. Osteosarcoma is one of the most common malignant tumors in adolescents, and research on its metastasis mechanisms and treatment effects faces the challenge that traditional models cannot provide an accurate physiological microenvironment; therefore, cell culture is necessary for such research.

[0003] Existing osteosarcoma cell culture methods employ independent culture, which involves single-stage culture with a single concentration of drug at the same time. To improve culture efficiency, multiple culture models and multiple control experiments are required to screen drug concentrations. However, these methods are insufficient in terms of material transport and mixing efficiency, and cannot achieve effective concentration gradient control, thus limiting their application in drug screening. Utility Model Content

[0004] This invention provides an in vitro culture chip for osteosarcoma, which solves the problem of low efficiency in drug concentration screening during in vitro culture of osteosarcoma.

[0005] To solve the above-mentioned technical problems, the technical solution adopted by this utility model is as follows: an osteosarcoma in vitro culture chip, including a carrier plate, with an outlet hole and at least two inlet holes at both ends of the carrier plate, the height of the inlet hole being higher than that of the outlet hole, and a mixing channel area, a concentration gradient forming area, a tissue culture area and a liquid storage buffer area arranged sequentially and in a gradually decreasing height between the inlet hole and the outlet hole. The concentration mixing channel area includes a connected mixing channel, an inflow end and an outflow end, with the inflow end and the outflow end respectively located on both sides of the mixing channel. Along the length of the mixing channel, each inflow end is located between adjacent outflow ends. The number of outflow ends in the concentration mixing channel area is one more than the number of inflow ends. The concentration gradient forming area consists of multiple areas arranged side by side, and also includes a flow channel. Each inlet hole is connected to each inflow end through the flow channel, and each outflow end is connected to each concentration gradient forming area through the flow channel.

[0006] In the preferred embodiment, the concentration mixing channel zone is multi-stage, with the outflow end of the upper-stage concentration mixing channel zone connected to the inflow end of the lower-stage concentration mixing channel zone, the inflow end of the uppermost concentration mixing channel zone connected to the inlet orifice, and the outflow end of the lowermost concentration mixing channel zone connected to the concentration gradient forming zone.

[0007] In the preferred embodiment, the flow channel is S-shaped.

[0008] In a preferred embodiment, a cover plate is also provided. One side of the cover plate is fastened to the carrier plate. The side of the cover plate that is fastened to the carrier plate is provided with a first baffle and a second baffle. The first baffle is provided at the end of the tissue culture zone near the concentration gradient formation zone to form a first narrowing channel. The second baffle is provided at the side of the liquid storage buffer zone near the tissue culture zone to form a second narrowing channel. Culture balls are provided in the tissue culture zone.

[0009] In the preferred embodiment, the carrier plate is further provided with a raised flange, and the cover plate is provided with a fastening flange, which abuts against the raised flange.

[0010] In a preferred embodiment, the cover plate has a perforated section, and a detachable inner plate frame is provided in the perforated section. The first and second stop bars are located on the buckle edge.

[0011] In the preferred embodiment, a transparent observation window is provided in the center of the inner panel frame.

[0012] In the preferred embodiment, multiple turbulence structures are provided within the flow channel.

[0013] The beneficial effects of this invention are as follows: It adopts a multi-entry pore combined with a mixing channel area and a concentration gradient forming area, which can effectively form a stable substance concentration gradient and provide a culture environment for cells that is closer to physiological conditions; it can simultaneously introduce multiple drugs of different concentrations into the same culture model and form a concentration gradient for the culture of osteosarcoma in the same tissue culture area, and can obtain multiple experimental results in the tissue culture area at the same time, with high experimental efficiency and intuitive and obvious comparison. Attached Figure Description

[0014] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0015] Figure 1 This is a simplified schematic diagram of a culture chip.

[0016] Figure 2 This is a schematic diagram of chip optimization.

[0017] Figure 3 This is an exploded view of the culture chip.

[0018] Figure 4 This is a schematic diagram of the mixing channel.

[0019] Figure 5 This is a schematic diagram of a pediatric detachable chip with an internal board frame.

[0020] Figure 6 This is a diagram showing the distribution of the flow channel turbulence structure.

[0021] Figure 7 This is a view of the bottom side of the cover plate.

[0022] Figure 8 This is a cross-sectional view of the chip's interior.

[0023] In the figure: cover plate 1; first baffle 101; second baffle 102; flange 103; carrier plate 2; inlet hole 201; flow channel 202; mixing channel 203; concentration gradient forming zone 204; separator 205; tissue culture zone 206; liquid storage buffer zone 207; outlet hole 208; raised flange 209; first sealing groove 210; inflow end 211; outflow end 212; turbulence structure 213; inner plate frame 3; observation window 4; culture ball 5; first narrowing channel 6; second narrowing channel 7. Detailed Implementation

[0024] Example 1:

[0025] like Figure 1-8 In an osteosarcoma in vitro culture chip, a carrier plate 2 is provided. The carrier plate 2 has an outlet hole 208 and at least two inlet holes 201 at its two ends. The inlet holes 201 are higher than the outlet holes 208. Between the inlet holes 201 and the outlet holes 208, there are sequentially connected and gradually decreasing height mixing channel region, concentration gradient forming region 204, tissue culture region 206, and liquid storage buffer 207. The concentration mixing channel region includes a connected mixing channel 203, an inflow end 211, and an outflow end 212. The inlet end 211 and the outlet end 212 are respectively located on both sides of the mixing channel 203. Along the length of the mixing channel 203, each inlet end 211 is located between adjacent outlet ends 212. The number of outlet ends 212 in the concentration mixing channel area is one more than the number of inlet ends 211. The concentration gradient forming area 204 consists of multiple areas arranged side by side and also includes a flow channel 202. Each inlet hole 201 is connected to each inlet end 211 through the flow channel 202, and each outlet end 212 is connected to each concentration gradient forming area 204 through the flow channel 202.

[0026] Separators 205 separate the concentration gradient formation zones 204, the tissue culture zone 206 is a single unit, and the liquid storage buffer zone 207 is a single unit.

[0027] Different concentrations of culture drugs flow in through two or more inlet holes 201, passing through the first horizontally placed mixing channel 203. Due to the narrowness of the mixing channel 203, the liquid mixing efficiency is low, thus exhibiting a concentration gradient distribution along the length of the mixing channel 203. Subsequently, the drugs flow into the next channel 203 from three or more flow channels 202, continuing to form four or more concentrations of culture medium, and finally flowing into the concentration gradient forming zone 204. After each drug of different concentrations is thoroughly mixed, it flows synchronously into the tissue culture zone 206.

[0028] In a preferred embodiment, the concentration mixing channel zone is multi-stage. The outflow end 212 of the upper-stage concentration mixing channel zone is connected to the inflow end 211 of the lower-stage concentration mixing channel zone. The inflow end 211 of the uppermost concentration mixing channel zone is connected to the inlet hole 201. The outflow end 212 of the lowermost concentration mixing channel zone is connected to the concentration gradient forming zone 204.

[0029] The upper and lower level regions are also connected through flow channel 202. The more levels passed through, the finer the gradient distribution.

[0030] In the preferred embodiment, the flow channel 202 is S-shaped.

[0031] S-shaped flow can increase the overall length of the flow channel, slow down the flow rate, increase the drug mixing time, and improve uniformity.

[0032] In a preferred embodiment, a cover plate 1 is also provided. One side of the cover plate 1 is fastened to the carrier plate 2. The side of the cover plate 1 that is fastened to the carrier plate 2 is provided with a first baffle 101 and a second baffle 102. The first baffle 101 is provided at one end of the tissue culture zone 206 near the concentration gradient forming zone 204 to form a first narrowing channel 6. The second baffle 102 is provided at one side of the liquid storage buffer zone 207 near the tissue culture zone 206 to form a second narrowing channel 7. A culture ball 5 is provided in the tissue culture zone 206.

[0033] Culture ball 5 is a solid sphere made of osteosarcoma cells and hydrogel, etc. It has a certain volume, which is larger than the width of the first narrowing channel 6 and the second narrowing channel 7, and is therefore confined in the tissue culture area 206.

[0034] In the preferred embodiment, the carrier plate 2 is further provided with a raised flange 209, and the cover plate 1 is provided with a fastening flange 103, which abuts against the raised flange 209.

[0035] Because the raised flange 209 is relatively high, the culture medium will not overflow even if the channel is blocked. The raised flange 209 can be provided with a first sealing groove 210 and a sealing strip can be placed on it to improve the sealing performance.

[0036] Furthermore, threaded holes can be machined on the raised flange 209, and through holes can be machined on the fastening flange 103, and the cover plate 1 and the carrier plate 2 can be locked together by bolts.

[0037] In the preferred embodiment, the cover plate 1 has a hollowed-out portion, and a detachable inner plate frame 3 is provided in the hollowed-out portion. The first stop bar 101 and the second stop bar 102 are provided on the buckle edge 103.

[0038] The outer side of the hollow section can be filled with a sealing strip using a sealing groove, and the inner plate frame 3 is installed with bolts and the sealing strip is pressed tightly.

[0039] In the preferred embodiment, the inner panel frame 3 has a transparent observation window 4 in the center.

[0040] The culture status of 206 species in the tissue culture area can be observed through observation window 4.

[0041] In a preferred embodiment, the flow channel 202 is provided with multiple flow disturbance structures 213.

[0042] The turbulence structure is a reverse herringbone structure, combined with an S-shaped concentration mixing channel, which increases the turbulence of the fluid, promotes the uniform mixing of different substances, and improves the material transport efficiency. It utilizes the principles of fluid mechanics to optimize the mixing effect by changing the channel shape and adding specific structures.

[0043] Example 2:

[0044] like Figure 1 A 3D-printed osteosarcoma in vitro culture chip includes a carrier chip and a cover plate. The carrier chip includes two inlets, a concentration mixing channel area, a concentration gradient forming area, a tissue culture area, an outlet, and a liquid storage area designed to prevent backflow and disruption of the concentration gradient. The concentration mixing channel area is provided with a concentration mixing channel. The concentration mixing channel area, concentration gradient forming area, tissue culture area, and liquid storage area are arranged in a stepped manner from high to low. The cover plate includes two baffles, which correspond to the tissue culture area and the liquid storage area of ​​the carrier chip, respectively.

[0045] Two liquids of different concentrations enter the concentration mixing channel zone through two inlets. After mixing, the liquid enters the concentration gradient forming zone. Once the concentration gradient is formed and stabilized, it flows into the tissue culture zone. The tumor spheres are placed in the tissue culture zone, allowing multiple tumor spheres to be evenly dispersed and simultaneously cultured at different drug concentrations on the same batch of tumor spheres. Finally, the liquid enters the liquid storage zone and flows out from the outlet.

[0046] The concentration mixing channel has at least three sections, with each channel in each section being S-shaped. The first section has two S-shaped channels, one end of which is connected to two inlets, and the other end is connected to the S-shaped channels in the next section via a transversely arranged channel. The number of channels increases by one with each subsequent section. This design ensures thorough and uniform mixing of the liquid within a limited area. The number and layers of S-shaped channels in the concentration mixing channel section can be adjusted according to experimental requirements to obtain different drug concentration gradients.

[0047] The concentration mixing channel is equipped with a reverse herringbone structure. Adding the reverse herringbone structure further ensures the uniformity of liquid mixing.

[0048] The raw materials for the carrier chip and cover plate include polylactic acid (PLA), which is produced using 3D printing technology. Using 3D printing to print the carrier chip and cover plate makes the external model of the main body more closely resemble actual needs, allowing for real-time adjustments and reprinting based on specific requirements. This method is fast and efficient; the PLA or resin materials used in printing are low-cost and non-toxic.

[0049] After the cover plate covers the carrier chip, it is fixed with bio-adhesive.

[0050] The above embodiments are merely preferred technical solutions of this utility model and should not be considered as limitations on this utility model. The protection scope of this utility model should be the technical solution described in the claims, including equivalent substitutions of the technical features described in the claims. That is, equivalent substitutions and improvements within this scope are also within the protection scope of this utility model.

Claims

1. An in vitro culture chip for osteosarcoma, characterized in that: The carrier plate (2) has an outlet hole (208) and at least two inlet holes (201) at both ends. The height of the inlet hole (201) is higher than that of the outlet hole (208). Between the inlet hole (201) and the outlet hole (208), there are interconnected and gradually decreasing height mixing channel area, concentration gradient forming area (204), tissue culture area (206) and liquid storage buffer area (207). The concentration mixing channel area includes interconnected mixing channel (203), inflow end (211) and outflow end (212). (212) are respectively set on both sides of the mixing channel (203). Each inflow end (211) is set between adjacent outflow ends (212) along the length of the mixing channel (203). The number of outflow ends (212) in the concentration mixing channel area is one more than the number of inflow ends (211). The concentration gradient forming area (204) consists of multiple side-by-side arrangements and also includes a flow channel (202). Each inlet hole (201) is connected to each inflow end (211) through the flow channel (202), and each outflow end (212) is connected to each concentration gradient forming area (204) through the flow channel (202).

2. The osteosarcoma in vitro culture chip according to claim 1, characterized in that: The concentration mixing channel zone is multi-level. The outflow end (212) of the upper-level concentration mixing channel zone is connected to the inflow end (211) of the lower-level concentration mixing channel zone. The inflow end (211) of the uppermost concentration mixing channel zone is connected to the inlet hole (201). The outflow end (212) of the lowermost concentration mixing channel zone is connected to the concentration gradient forming zone (204).

3. The osteosarcoma in vitro culture chip according to claim 1, characterized in that: The flow channel (202) is S-shaped.

4. The osteosarcoma in vitro culture chip according to claim 1, characterized in that: A cover plate (1) is also provided. One side of the cover plate (1) is fastened to the carrier plate (2). The side of the cover plate (1) that is fastened to the carrier plate (2) is provided with a first baffle (101) and a second baffle (102). The first baffle (101) is located at one end of the tissue culture area (206) near the concentration gradient formation area (204) to form a first narrowing channel (6). The second baffle (102) is located at one side of the liquid storage buffer zone (207) near the tissue culture area (206) to form a second narrowing channel (7). A culture ball (5) is provided in the tissue culture area (206).

5. The osteosarcoma in vitro culture chip according to claim 1, characterized in that: The carrier plate (2) is also provided with a raised flange (209), and the cover plate (1) is provided with a fastening flange (103), which is attached to the raised flange (209).

6. The osteosarcoma in vitro culture chip according to claim 4, characterized in that: The cover plate (1) has a hollowed-out part, and a detachable inner plate frame (3) is provided in the hollowed-out part. The first stop bar (101) and the second stop bar (102) are provided on the buckle edge (103).

7. The osteosarcoma in vitro culture chip according to claim 6, characterized in that: The inner panel (3) has a transparent observation window (4) in the center.

8. The osteosarcoma in vitro culture chip according to claim 1, characterized in that: Multiple turbulence structures (213) are provided inside the flow channel (202).

Images