Organoid culture medium exchanger

Abstract

This utility model discloses an organoid culture medium exchanger, comprising: a pipette tip assembly, including: a pipette tip, a filter, and a suction nozzle, wherein the pipette tip, filter, and suction nozzle are all conical in shape; the filter includes a first end and a second end, and the diameter of the filter gradually increases from the first end to the second end; the first end of the filter is inserted into the pipette tip, and the suction nozzle is inserted into the second end of the filter; a culture medium recovery bottle; an air pump; a first conduit, one end of which is connected to the suction nozzle, and the other end of which is inserted into the culture medium recovery bottle; and a second conduit, one end of which is connected to the air pump, and the other end of which is inserted into the culture medium recovery bottle; wherein the culture medium recovery bottle is provided with a one-way inlet valve and a one-way outlet valve at the bottle opening; the first conduit is inserted into the culture medium recovery bottle through the one-way inlet valve, and the second conduit is inserted into the culture medium recovery bottle through the one-way outlet valve. This application allows for the continuous aspiration and storage of waste liquid, and is also applicable to micro-organoids.

Description

Technical Field

[0001] This utility model relates to the field of in vitro culture of organoids, and in particular to an organoid culture medium exchanger. Background Technology

[0002] Organoids are tissue analogs with a certain spatial structure formed by in vitro three-dimensional (3D) culture of adult stem cells or pluripotent stem cells. At different stages of organoid culture, the culture medium often needs to be changed every few days due to the continuous metabolism of cells during the culture process. Current pipettes have a limited capacity for removing liquid at a time, making continuous aspiration of waste liquid impossible. Changing the medium for organoids cultured in multi-well plates requires repeated aspiration and drainage operations, which is cumbersome, time-consuming, and labor-intensive. Since organoid formation requires 3D suspension culture and the formed organoids are relatively small, using a pipette for medium changes can easily result in the direct aspiration of these small organoids, causing experimental losses. Furthermore, the suction force of the pipette tip in current technology is uncontrollable, which may cause direct damage to some organoids (such as whole-brain organoids) due to improper handling. Utility Model Content

[0003] The main objective of this invention is to provide an organoid culture medium exchanger to solve the problems of existing exchangers being unable to continuously remove waste liquid and being unsuitable for small organoids.

[0004] According to an embodiment of this utility model, an organoid culture medium exchanger is provided, comprising: a pipette tip assembly, the pipette tip assembly including: a pipette tip, a filter, and a suction nozzle, wherein the pipette tip, the filter, and the suction nozzle are all conical in shape, the filter including a first end and a second end, the diameter of the filter gradually increasing from the first end towards the second end, the first end of the filter being inserted into the pipette tip and tightly connected to the pipette tip, and the suction nozzle being inserted into the second end of the filter and tightly connected to the second end of the filter; a culture medium recovery bottle; an air pump; a first conduit, one end of the first conduit communicating with the suction nozzle, and the other end inserted into the culture medium recovery bottle; a second conduit, one end of the second conduit communicating with the air pump, and the other end inserted into the culture medium recovery bottle; wherein the culture medium recovery bottle is provided with a one-way inlet valve and a one-way outlet valve at the bottle opening, the first conduit being inserted into the culture medium recovery bottle through the one-way inlet valve, and the second conduit being inserted into the culture medium recovery bottle through the one-way outlet valve.

[0005] The filter includes a metal filter screen with a pore size of 0.05 mm.

[0006] The filter screen is disposed in the filter at its first end near the filter.

[0007] The first conduit includes a first tube section and a second tube section. The first tube section is connected to the suction nozzle and the one-way inlet valve. The second tube section is inserted into the culture medium recovery bottle through the one-way inlet valve and is located at the bottle opening of the culture medium recovery bottle.

[0008] The second conduit includes a first tube section and a second tube section. The first tube section is connected to the one-way vent valve and the air pump. The second tube section is inserted into the culture medium recovery bottle through the one-way vent valve and is located at the bottle opening of the culture medium recovery bottle.

[0009] Wherein, the length of the second segment of the first catheter is greater than the length of the second segment of the second catheter.

[0010] The organoid culture medium exchanger further includes a base, in which the air pump is disposed, and the base includes an upward-opening cavity, in which the culture medium recovery bottle is inserted.

[0011] The base also includes a support frame, and the suction nozzle is fitted into the support frame.

[0012] The nozzle is provided with a sealing mechanism, and a touch-sensitive physical switch for controlling the opening and closing of the sealing mechanism is provided on the side surface of the nozzle.

[0013] The first catheter and the second catheter are flexible catheters.

[0014] According to the technical solution of this utility model, by setting a filter between the pipette tip and the nozzle, and connecting the nozzle to the culture medium recovery bottle through a first conduit and a one-way liquid inlet valve, and connecting the air pump to the culture medium recovery bottle through a second conduit and a one-way air outlet valve, not only is the function of continuously aspirating waste liquid realized, but the aspirated waste liquid is also temporarily stored in the culture medium recovery bottle, which improves the efficiency of the medium replacement operation. It also prevents small organoids from being aspirated along with the culture medium, reducing sample loss. Attached Figure Description

[0015] The accompanying drawings, which are included to provide a further understanding of the present invention and form part of this application, illustrate exemplary embodiments of the present invention and, together with the description thereof, serve to explain the present invention and do not constitute an undue limitation thereof. In the drawings:

[0016] Figure 1 This is a schematic diagram of the structure of an organoid culture medium exchanger according to an embodiment of the present invention;

[0017] Figure 2 This is a schematic diagram of the gun head assembly according to an embodiment of the present utility model;

[0018] Figure 3 This is a schematic diagram of the filter structure according to an embodiment of the present utility model;

[0019] Figure 4 yes Figure 2 A cross-sectional view of line AA in the middle. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of this utility model clearer, the technical solutions of this utility model will be clearly and completely described below in conjunction with specific embodiments and corresponding drawings. Obviously, the described embodiments are only a part of the embodiments of this utility model, and not all of them. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this utility model.

[0021] The technical solutions provided by the various embodiments of this utility model are described in detail below with reference to the accompanying drawings.

[0022] According to an embodiment of this utility model, an organoid culture medium exchanger is provided, which is applicable to the culture medium exchange of organoids such as whole-brain organoids, intestinal organoids, and lung organoids. (Reference) Figure 1 The organoid culture medium exchanger includes: a pipette tip assembly, a culture medium recovery bottle 4, a first conduit, a second conduit, an air pump 61, and a sealing valve 8 located at the mouth of the culture medium recovery bottle 4. The first conduit, the second conduit, and the culture medium recovery bottle 4 constitute a culture flask (system). The pipette tip assembly is connected to the culture medium recovery bottle 4 via the first conduit and the sealing valve 8. The culture medium recovery bottle 4 is connected to the air pump 61 via the sealing valve 8 and the second conduit. The air pump 61 draws air from the culture medium recovery bottle 4, creating a stable negative pressure environment within the culture flask (system). The pipette tip assembly draws the organoid culture medium into the culture medium recovery bottle 4, thereby achieving continuous aspiration and temporary storage of the culture medium, improving exchange efficiency, and providing convenience and speed.

[0023] Reference Figures 1-3The pipette tip assembly includes: a pipette tip 1, a filter 2, and a pipette nozzle 3. The pipette tip 1 is conical in shape, and its capacity can be selected according to the size of the organoid culture well, typically 20-100 μl. The pipette tip 1 can be made of medical-grade silicone. The filter 2 is also conical in shape, with its small-diameter end (first end) inserted into and tightly fitted to the large-diameter end of the pipette tip 1 to avoid gaps. A filter screen 21 is disposed inside the filter 2, positioned near the small-diameter end of the filter 2. The filter screen 21 can be a metal filter, and its pore size is 0.05 mm. The filter screen 21 prevents the aspiration of small organoids (organoid clusters with a diameter less than 200 μm), thereby reducing experimental losses. The pipette nozzle 3 is also conical in shape, and its material can be medical-grade silicone. The small-diameter end of the pipette nozzle 3 is inserted into and tightly fitted to the large-diameter end of the filter 2 to avoid gaps. A switch 31 (physical switch) is provided on the side surface of the pipette nozzle 3. The switch 31 can be a touch-sensitive push switch. A sealing mechanism controlled by the switch 31 can be provided in the pipette nozzle 3. By opening or closing the sealing mechanism, the opening or closing of the pipette nozzle 3 can be controlled, thereby controlling whether the culture flask (system) is connected to the outside. In the embodiment of this application, through holes are provided near the large-diameter end and the small-diameter end of the pipette nozzle 3, respectively. The first conduit is inserted into the pipette nozzle 3 through the through hole at the large-diameter end of the pipette nozzle 3, and after passing through the sealing mechanism in the pipette nozzle 3, the first conduit exits from the through hole at the small-diameter end of the pipette nozzle 3, so that the first conduit is connected to the pipette head 1 and the filter 2 through the pipette nozzle 3. Specifically, refer to Figure 4The sealing mechanism includes a first sealing unit 32, a second sealing unit 33, and springs 34. The first sealing unit 32 is connected to a switch 31. Pressing the switch 31 moves the first sealing unit 32 within the suction nozzle 3 in the direction of the switch 31's press. A protrusion is provided on the first sealing unit 32, the height of which matches the height of the first guide tube in the suction nozzle 3. The second sealing unit 33 is a fixed structure with a groove for accommodating the first guide tube. The position and shape of the groove in the second sealing unit 33 match the position and shape of the protrusion in the first sealing unit 32. Multiple springs 34 are arranged between the first and second sealing units 32, allowing the user to control the pressure exerted by pressing the switch 31 to adjust the degree of compression of the first guide tube by the protrusion, thereby changing the degree of sealing and controlling the suction force of the nozzle assembly. When the user releases the switch 31, the first sealing unit 32 returns to its original position via the springs 34. The protrusion and groove can be semi-cylindrical or other shapes. The sealing mechanism can be closed by applying a moderate force (approximately 1-2 N) to switch 31, allowing the user to control the gas pressure inside the culture flask (system) with one hand while holding the pipette tip assembly. When the sealing mechanism is open, the culture flask (system) is connected to the outside environment, and the organoid culture medium exchanger can draw culture medium. When the sealing mechanism is closed, the culture flask (system) is in a temporarily closed state, and the organoid culture medium exchanger stops drawing culture medium, thus achieving efficient, precise, and immediate control of the exchanger's use. The pipette tip 1 and filter 2 can be disposable or reusable. When the pipette tip 1 and filter 2 are used multiple times, they can be sterilized by high temperature or alcohol immersion.

[0024] The culture medium recovery bottle 4 can be a standard laboratory glass solution bottle, the volume of which can be selected according to the volume of liquid to be aspirated, preferably 1L. A sealing valve 8 is installed at the mouth of the culture medium recovery bottle 4, and the sealing valve 8 is sealed to the culture medium recovery bottle 4. The sealing valve 8 is equipped with two one-way valves, namely a one-way liquid inlet valve 81 and a one-way gas outlet valve 82. The one-way liquid inlet valve 81 only allows external liquid to enter the culture medium recovery bottle 4 through it, and the one-way gas outlet valve 82 only allows gas in the culture medium recovery bottle 4 to exit through it. The pipette 3 is connected to the culture medium recovery bottle 4 through a first conduit and the one-way liquid inlet valve 81. Specifically, the first conduit is divided into two sections, 51 and 52, by the liquid inlet valve 81, with the length of section 51 being much greater than the length of section 52. Section 51 is a soft or flexible conduit, and the section 51 connects the large-diameter end of the pipette 3 to the end of the one-way liquid inlet valve 81 located on the outside of the culture medium recovery bottle 4. The tube body 52 can be a flexible, soft, or rigid conduit. The tube body 52 is inserted into one end of the one-way inlet valve 81 located inside the culture medium recovery bottle 4, and is positioned at the bottle opening of the culture medium recovery bottle 4. In this embodiment, when the tube body 51 and tube body 52 are flexible or soft conduits, they can be made of rubber tubing.

[0025] According to an embodiment of this application, the fluid exchanger further includes a base 6, which includes an open cavity with its opening facing upwards. The culture medium recovery bottle 4 can be inserted into the cavity of the base 6. This not only reduces the volume and floor space of the organoid culture fluid exchanger but also prevents the culture medium recovery bottle 4 from tipping over due to accidental contact. Below the cavity of the base 6, there is space where an air pump 61 can be installed. The air pump 61, also known as a vacuum pump or air pump, has a negative pressure generation time of ≤0.5 seconds to ensure real-time controllable fluid extraction. Furthermore, a battery powering the air pump 61 can also be installed in this space.

[0026] A switch 62 is also provided on the side wall of the base 6. Switch 62 is equivalent to the main switch of this type of organ culture medium exchanger. It can be used to control the power supply of the air pump 61. When switch 62 is closed, the battery supplies power to the air pump 61, and the air pump 61 starts to work. A support frame 63 is also provided on the base 6. The height of the support frame 63 is greater than the height of the bottle mouth of the culture medium recovery bottle 4. When the pipette tip assembly is not in use, the nozzle 3 can be locked in the support frame 63, thereby preventing any liquid that may remain in the tube 51 from flowing out through the pipette tip assembly.

[0027] The culture medium recovery bottle 4 is connected to the air pump 61 via a one-way vent valve 82 and a second conduit. Specifically, the second conduit is divided into two sections, 71 and 72, by the vent valve 82, with section 71 being significantly longer than section 72. Section 71 is a flexible or soft conduit, and it connects to the air pump 61 and the one-way vent valve 82 at the outer end of the culture medium recovery bottle 4. Section 72 can be a flexible, soft, or rigid conduit, and it is inserted into the one-way vent valve 82 at the inner end of the culture medium recovery bottle 4, with section 72 positioned at the bottle opening. In this embodiment, when sections 71 and 72 are flexible or soft conduits, they can be made of rubber tubing. Furthermore, by positioning tubes 52 and 72 at the mouth of the culture medium recovery bottle 4, the culture medium already collected in the culture medium recovery bottle 4 can be prevented from contacting the tubes located in the culture medium recovery bottle 4; and the length of tube 52 is longer than the length of tube 72, that is, the liquid outlet end of tube 52 is lower than the air inlet end of tube 72, so as to prevent the culture medium flowing out through tube 52 from being accidentally sucked into tube 72 due to excessive suction of air pump 61.

[0028] In actual operation, firstly, select a pipette tip 1 of appropriate size and assemble the pipette tip 1 and filter 2 onto the pipette nozzle 3 to form a pipette tip assembly; secondly, turn on the switch 62 on the base 6, the air pump 61 starts working, a negative pressure is formed inside the culture bottle (system), the two one-way valves in the sealing valve 8 open, and the space inside the culture bottle (system) is connected to the outside; then, insert the pipette tip assembly vertically about 5mm below the surface of the culture medium to draw the culture medium, the air in the culture medium recovery bottle 4 is drawn away through the one-way air outlet valve 82, and the culture medium is drawn by the pipette tip 1, filtered by the filter 2, and enters the culture medium recovery bottle 4 through the pipette nozzle 3, the first conduit and the one-way inlet valve 81.

[0029] After the culture medium in a single culture well is aspirated, press switch 31 to temporarily stop the liquid exchanger from aspirating. At this time, the pipette tip assembly can be inserted into the next culture well to be aspirated. Release switch 31 and repeat the above steps to continue aspirating the culture medium until all the culture wells to be aspirated have been aspirated. Finally, turn off switch 62, and the air pressure in the culture medium recovery bottle 4 will return to normal pressure. The one-way liquid inlet valve 81 and the one-way air outlet valve 82 will automatically close.

[0030] After aspirating the culture medium from all wells, pipette tip 1 and filter 2 can be removed and sterilized using high-temperature autoclaving (121°C, 15 minutes) or by soaking pipette tip 1, filter 2, or other structures that came into contact with the culture medium in 75% alcohol to avoid unsanitary conditions. Waste liquid in culture medium recovery bottle 4 can also be collected and disposed of at this time.

[0031] This utility model's organoid culture medium exchanger is suitable for high-hygiene environments such as organoid culture and stem cell research, and has broad market prospects. It has at least one of the following beneficial effects:

[0032] 1. The integrated pipette tip assembly design is convenient to operate. Liquid can be aspirated by operating the pipette tip assembly with one hand, which is highly flexible in operation. It can also continuously aspirate culture medium waste liquid and collect and store it for subsequent processing. Compared with traditional pipettes, which require aspirating and releasing the culture medium before the next aspiration, the operating efficiency of this type of organ culture medium exchanger is significantly improved.

[0033] 2. Install a filter between the pipette tip and the nozzle to protect the cultured organoids, prevent the small organoids from being aspirated along with the culture medium, and reduce experimental losses.

[0034] 3. With fewer pipe interfaces and a compact structure, all components in contact with the culture medium are detachable for sterilization, preventing bacterial growth and contamination of organoids, thus improving the hygienic performance of the fluid exchanger.

[0035] 4. Small size: The volume of this type of organ culture medium exchanger is 1 / 3 of that of a traditional multi-channel exchanger, making it more suitable for use in space-constrained clean benches.

[0036] 5. A switch is provided on the nozzle, allowing for one-handed control of the suction power of the nozzle assembly via a physical switch. The suction power is controllable and the operation is simple.

[0037] Although this disclosure has been described in detail with reference to specific embodiments thereof, those skilled in the art will understand that various changes and modifications may be made therein without departing from the spirit and scope of the embodiments. Therefore, this disclosure is intended to cover modifications and variations thereof, and any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this invention should be included within the scope of the claims of this disclosure and their equivalents.

[0038] Furthermore, features disclosed in the foregoing description, claims, or drawings, expressed in their specific form or according to the manner of performing the disclosed function or the method or process for obtaining the disclosed result, may, as appropriate, be used alone or in any combination of these features to implement the present invention in their different forms. Specifically, one or more features of any embodiment described herein may be combined with one or more features of any other embodiment described herein.

[0039] Protection may also be sought for any features disclosed in any one or more public documents combined with this disclosure and / or merged by reference.

Claims

1. A medium exchanger for organoid culture, characterized in that, include: A nozzle assembly includes a suction head, a filter, and a suction nozzle, wherein the suction head, the filter, and the suction nozzle are all conical in shape, the filter includes a first end and a second end, the diameter of the filter gradually increases from the first end to the second end, the first end of the filter is inserted into the suction head and tightly connected to the suction head, and the suction nozzle is inserted into the second end of the filter and tightly connected to the second end of the filter; Culture medium recovery bottle; air pump; A first catheter, one end of which is connected to the suction nozzle, and the other end of which is inserted into the culture medium recovery bottle; The second conduit has one end connected to the air pump and the other end inserted into the culture medium recovery bottle; The culture medium recovery bottle is equipped with a one-way inlet valve and a one-way outlet valve at its opening. The first conduit is inserted into the culture medium recovery bottle through the one-way inlet valve, and the second conduit is inserted into the culture medium recovery bottle through the one-way outlet valve.

2. The organoid culture medium exchanger according to claim 1, characterized in that, The filter is equipped with a metal filter screen with a pore size of 0.05 mm.

3. The organoid culture medium exchanger according to claim 2, characterized in that, The filter screen is disposed in the filter near the first end of the filter.

4. The organoid culture medium exchanger according to claim 1, characterized in that, The first catheter includes a first tube section and a second tube section. The first tube section is connected to the suction nozzle and the one-way inlet valve. The second tube section is inserted into the culture medium recovery bottle through the one-way inlet valve and is located at the bottle opening of the culture medium recovery bottle.

5. The organoid culture medium exchanger according to claim 4, characterized in that, The second conduit includes a first tube section and a second tube section. The first tube section is connected to the one-way vent valve and the air pump. The second tube section is inserted into the culture medium recovery bottle through the one-way vent valve and is located at the bottle opening of the culture medium recovery bottle.

6. The organoid culture medium exchanger according to claim 5, characterized in that, The length of the second segment of the first catheter is greater than the length of the second segment of the second catheter.

7. The organoid culture medium exchanger according to claim 1, characterized in that, Also includes: The base, in which the air pump is disposed, includes an upward-opening cavity, and the culture medium recovery bottle is inserted into the cavity.

8. The organoid culture medium exchanger according to claim 7, characterized in that, The base also includes a support frame, and the suction nozzle is fitted in the support frame.

9. The organoid culture medium exchanger according to claim 1, characterized in that, The nozzle is provided with a sealing mechanism, and a touch-sensitive physical switch for controlling the opening and closing of the sealing mechanism is provided on the side surface of the nozzle.

10. The organoid culture medium exchanger according to claim 1, characterized in that, The first catheter and the second catheter are flexible catheters.

Images