Mesenchymal stem cell separation device

Abstract

The utility model relates to a mesenchymal stem cell separation device, which comprises a dish body, a dish cover and a liquid mechanism, the periphery of the bottom of the dish body extends downwards to form a lower edge, the bottom surface of the dish body sinks inwards to form a plurality of ridges, two sides of each ridge are abrupt slope surfaces, and the ridges are radially and circumferentially arranged on the bottom surface of the dish body. A V-shaped liquid permeable groove parallel to the ridges on the two sides is formed between every two adjacent ridges on the bottom surface of the dish body; the liquid injection mechanism comprises a liquid injection cavity with the lower portion closed up and a liquid permeating cavity with the upper portion unfolded, a liquid inlet connector and a liquid outlet connector are arranged on the liquid injection cavity, and a liquid injection pipe and a liquid outlet pipe are arranged on the lower portion of the liquid injection cavity. According to the mesenchymal stem cell separation device, the mode of ascending infiltration and descending emptying at the bottom is achieved, a culture solution is updated on the premise that the dish cover does not need to be opened and closed, meanwhile, a stem cell culture space is isolated from a liquid injection space and a liquid drainage space, damage to stem cells caused by injection and drainage of the culture solution is prevented, the dish cover does not need to be opened and closed in the culture solution updating process, and the culture solution updating efficiency is improved. The probability that the separated stem cells are polluted is effectively reduced.

Description

Technical Field

[0001] The utility model relates to the field of biotechnology and equipment, and in particular to a culture device for separating and purifying mesenchymal stem cells. Background Art

[0002] Currently, the most common method for culturing and isolating mesenchymal stem cells is tissue adherence culture. This involves removing blood vessels and clots, then mincing and washing the tissue. The tissue is then cultured in a culture dish, where the stem cells adhere to the dish, grow, and proliferate, forming clusters that can then be isolated. Stem cell growth produces a large amount of metabolic waste that enters the culture medium. A certain accumulation of metabolites can slow or even hinder stem cell growth and clustering, necessitating frequent culture medium replenishment. This process requires repeated handling of the culture dish, opening and closing the lid, and pipetting and injecting culture medium. This not only easily contaminates the sample, but also, in the early stages of stem cell adherence, the small size of the clusters can easily be disrupted by the fluid being drawn in and out of the cell clusters. Utility Model Content

[0003] The technical problem to be solved by the present invention is to overcome the defects of the existing method of updating the culture medium during the proliferation culture process after stem cell culture and separation, which causes sample contamination and damage to adherent cell clusters. A mesenchymal stem cell separation device is provided. The device, through the dish body structure design, realizes the method of increasing infiltration and decreasing emptying of the culture medium by filling and discharging the culture medium at the bottom, and updates the culture medium without opening and closing the dish cover. At the same time, the stem cell culture space is isolated from the injection and discharge spaces, preventing damage to the stem cells by the injection and discharge of the culture medium. The culture medium renewal process does not require opening and closing the dish cover, effectively reducing the probability of contamination of the separated stem cells.

[0004] The mesenchymal stem cell separation device includes a glass dish body and a dish cover for covering the open top of the dish body, and also includes a liquid injection mechanism arranged below the dish body, wherein the outer periphery of the bottom of the dish body extends downward to form a lower edge, and the bottom surface of the dish body is recessed inward to form a plurality of ridges with steep slopes on both sides, the ridges extend outward from the center of the bottom surface of the dish body, and the ridges are radially arranged circumferentially on the bottom surface of the dish body, and a V-shaped liquid permeable groove parallel to the ridges on both sides is opened between two adjacent ridges on the bottom surface of the dish body; a section of inward-recessed thread is formed on the upper part of the inner wall of the lower edge of the dish body; the liquid injection mechanism is funnel-shaped, including a liquid injection cavity that converges at the bottom and a liquid permeable cavity that expands at the upper part, and the liquid injection cavity is provided with a liquid inlet interface and a liquid discharge interface, and the lower part of the liquid injection cavity is provided with an injection tube and a liquid discharge tube, the inner end of which is connected to the liquid injection cavity and the outer end extends outward to the outside of the liquid permeable cavity shielding area; the outer wall of the liquid permeable cavity is provided with screw threads matching the screw threads.

[0005] Furthermore, a partition extending from the upper edge of the liquid injection cavity to the bottom surface of the liquid permeable cavity is provided in the liquid injection mechanism cavity, the lower part of the partition is located between the liquid injection pipe and the liquid discharge pipe, and a gap is provided at the rear of the partition.

[0006] Furthermore, both the injection pipe and the discharge pipe are provided with a stop valve.

[0007] Furthermore, a sealing rubber ring is provided on the upper inner portion of the lower edge.

[0008] Furthermore, an open groove is provided at the lower portion of the lower edge.

[0009] The utility model discloses a mesenchymal stem cell separation device, which overcomes the defects of the existing stem cell culture and separation proliferation culture process caused by the culture medium renewal method, that is, easy contamination of samples and damage to adherent cell clusters. The device realizes the method of rising infiltration and descending emptying of the culture medium by filling and discharging the culture medium at the bottom through the dish body structure design, and the culture medium is renewed without opening and closing the dish cover. At the same time, the stem cell culture space is isolated from the injection and discharge spaces, preventing the culture medium from being damaged by the injection and discharge. The culture medium renewal process does not require opening and closing the dish cover, which effectively reduces the probability of contamination of the separated stem cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The following is a further description of the mesenchymal stem cell separation device of the present invention in conjunction with the accompanying drawings:

[0011] Figure 1 This is a schematic diagram of the planar structure of the mesenchymal stem cell separation device;

[0012] Figure 2 This is a schematic diagram of the three-dimensional structure of the mesenchymal stem cell separation device;

[0013] Figure 3 yes Figure 2 Schematic diagram of the structure viewed from above;

[0014] Figure 4 yes Figure 1 AA section view;

[0015] Figure 5 yes Figure 2 Schematic diagram of the structural explosion;

[0016] Figure 6 yes Figure 3 Schematic diagram of the structural explosion.

[0017] In the picture:

[0018] 1-dish body; 11-lower edge, 12-ridge, 13-V-shaped liquid permeable groove, 14-sealing rubber ring; 101-thread, 111-opening groove;

[0019] 2-dish cover;

[0020] 3-liquid injection mechanism; 31-liquid injection chamber, 32-liquid permeable chamber, 33-partition, 34-stop valve; 301-screw thread, 311-liquid injection pipe, 312-liquid discharge pipe, 331-notch. DETAILED DESCRIPTION

[0021] In this utility model, unless otherwise specified or limited, the terms "installed," "connected," "connect," "fixed," etc. should be understood in a broad sense. For example, they can refer to fixed connection, detachable connection, or integration; they can refer to direct connection or indirect connection through an intermediate medium; they can refer to internal communication between two components or interaction between two components. Those skilled in the art will understand the specific meanings of the above terms in this utility model based on specific circumstances.

[0022] In the description of the present invention, it should be understood that the terms "left", "right", "front", "back", "top", "bottom", "inside", "outside", etc., indicating directions or positional relationships, are based on the directions or positional relationships shown in the accompanying drawings. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction. Therefore, they cannot be understood as limitations on the present invention.

[0023] The technical solution of the present invention is further described below with specific embodiments, but the protection scope of the present invention is not limited to the following embodiments.

[0024] Implementation method 1: Figures 1 to 6 As shown, the mesenchymal stem cell separation device includes a glass dish 1 and a dish cover 2 for covering the upper opening of the dish 1, and further includes a liquid injection mechanism 3 arranged below the dish 1, wherein the bottom periphery of the dish 1 extends downward to form a lower edge 11, and the bottom surface of the dish 1 is inwardly recessed to form a plurality of ridges 12 with steep slopes on both sides, the ridges 12 extending outward from the center of the bottom surface of the dish 1, and the ridges 12 are radially arranged circumferentially on the bottom surface of the dish 1, and the bottom surface of the dish 1 is provided with a ridge 12 between two adjacent ridges 12. Parallel V-shaped liquid permeable grooves 13; a section of inward-sunken thread 101 is formed on the upper part of the inner wall of the lower edge of the dish body 1; the liquid injection mechanism 3 is funnel-shaped, including a liquid injection cavity 31 that converges at the bottom and a liquid permeable cavity 32 that expands at the top, and the liquid injection cavity is provided with a liquid inlet interface and a liquid discharge interface. The lower part of the liquid injection cavity 31 is provided with a liquid injection tube 311 and a liquid discharge tube 312, the inner end of which is connected to the liquid injection cavity 31 and the outer end extends outside the shielding area of ​​the liquid permeable cavity 32; the outer wall of the liquid permeable cavity 32 is provided with a screw thread 301 that matches the thread.

[0025] Embodiment 2: The injection mechanism 3 of this mesenchymal stem cell culture device is equipped with a partition 33 extending from the top edge of the injection chamber 31 to the bottom surface of the permeable chamber 32. The lower portion of the partition 33 is located between the injection tube 311 and the drainage tube 312. A notch 331 is provided at the rear of the partition 33. This partition circulates cleaning fluid during flushing. Cleaning fluid is injected from the injection tube and flows through the notch to the drainage port. This prevents the newly injected cleaning fluid from flowing directly into the drainage tube and being discharged, preventing the stem cells in the dish from being fully cleaned. The remaining structure and components are as described in Embodiment 1 and will not be repeated here.

[0026] Embodiment 3: Both the injection tube 311 and the discharge tube 312 of this mesenchymal stem cell culture device are equipped with on / off valves 34. When connected to an automated injection device via the catheter, these on / off valves can be electromagnetic on / off valves or variable-speed valves for automatic control. When manually injecting or discharging liquids, these on / off valves can be variable-speed valves for flow rate control and reduced errors. The remaining structure and components are as described in Embodiment 1 and will not be repeated here.

[0027] Implementation 4: A sealing rubber ring 14 is provided on the upper portion of the dish holder 11 of the mesenchymal stem cell culture device. This sealing rubber ring is used to provide an air-liquid seal between the upper edge of the sealing rubber ring and the bottom surface of the dish body after the liquid injection mechanism is threadedly connected to the dish holder, thereby preventing contamination and leakage. An open groove 111 is provided on the lower portion of the dish holder 11 of the mesenchymal stem cell culture device. This notch exposes the inlet and outlet pipe areas after the liquid injection mechanism is threadedly connected to the dish holder, facilitating the connection of a catheter, syringe, or other liquid injection and drainage equipment. The remaining structures and components are as described in Implementation 1 and will not be repeated.

[0028] When in use: Lay the cell isolation filter cloth on the lower edge of the clean bench and press it against the bottom of the dish with a sealing rubber ring. Then connect the injection mechanism to the bottom of the lower edge and tighten it. After injecting enough culture fluid through the injection tube, place the chopped tissue in. Cover the dish with the lid and drain part of the culture fluid by gravity through the drainage tube to make the cell clusters sink and adhere to the wall. The ridges in the dish increase the area of ​​the vertical wall so that stem cells can grow and eventually form large cell clusters. The sides of the ridges are inclined to facilitate the final cell clusters to be flushed down and recovered. The culture fluid is replaced by operating the valves of the injection tube and the drainage tube. The injected liquid is The narrow gap formed by the permeable cavity and the bottom of the dish allows liquid to penetrate into the dish through the filter cloth and V-shaped permeable groove, infiltrating the stem cell clusters. The permeable position of the V-shaped permeable groove is close to but slightly away from the ridge used to grow the cell clusters, ensuring that liquid does not directly flush the sides of the ridge when it is injected. Without opening the lid, the pipette injection pressure is reduced, which could cause the stem cells to be washed off the wall in the early stages of clustering and cause cell damage. The expanded and flattened permeable cavity above the funnel-shaped injection mechanism saves on the amount of reagent culture medium used. The stem cells inside the dish are isolated throughout the process, effectively preventing contamination of the stem cells separated by adherence to the wall.

[0029] This mesenchymal stem cell separation device overcomes the defects of easy sample contamination and damage to adherent cell clusters caused by the culture medium renewal method during the proliferation culture process after stem cell culture separation. Through the dish body structure design, it realizes the method of rising infiltration and descending emptying of the culture medium injection and discharge at the bottom, and the culture medium is updated without opening and closing the dish cover. At the same time, the stem cell culture space is isolated from the injection and discharge spaces to prevent the damage of the culture medium injection and discharge to the stem cells. The culture medium renewal process does not require opening and closing the dish cover, which effectively reduces the probability of contamination of the separated stem cells.

[0030] The above description shows the main features, basic principles, and advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments or examples, and that the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, the above embodiments or examples should be regarded as exemplary and non-restrictive. The scope of the present invention is limited by the appended claims rather than the above description, and it is intended that all changes that fall within the meaning and scope of the equivalent elements of the claims are included in the present invention. Any figure mark in the claims should not be regarded as limiting the claims involved.

[0031] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A mesenchymal stem cell separation device, characterized by: The invention comprises a glass dish body (1) and a dish cover (2) for covering the upper opening of the dish body (1), and further comprises a liquid injection mechanism (3) arranged below the dish body (1), wherein: The bottom periphery of the dish body (1) extends downward to form a lower edge (11), and the bottom surface of the dish body (1) is recessed inward to form a plurality of ridges (12) with steep slopes on both sides. The ridges (12) extend outward from the center of the bottom surface of the dish body (1), and the ridges (12) are arranged radially around the bottom surface of the dish body (1). The bottom surface of the dish body (1) is provided with a V-shaped liquid-permeable groove (13) parallel to the ridges (12) on both sides between two adjacent ridges (12); the upper part of the inner wall of the lower edge of the dish body (1) is formed with a recessed The liquid injection mechanism (3) is funnel-shaped and comprises a liquid injection cavity (31) that converges at the bottom and a liquid permeable cavity (32) that expands at the top. The liquid injection cavity is provided with a liquid inlet interface and a liquid discharge interface. The lower part of the liquid injection cavity (31) is provided with a liquid injection tube (311) and a liquid discharge tube (312), the inner end of which is connected to the liquid injection cavity (31) and the outer end of which extends outside the shielding area of ​​the liquid permeable cavity (32). The outer wall of the liquid permeable cavity (32) is provided with screw threads (301) that match the screw threads.

2. The mesenchymal stem cell separation device according to claim 1, wherein: A partition (33) is provided in the liquid injection mechanism (3) and extends from the upper edge of the liquid injection cavity (31) to the bottom surface of the liquid permeable cavity (32). The lower portion of the partition (33) is located between the liquid injection pipe (311) and the liquid discharge pipe (312). A notch (331) is provided at the rear of the partition (33).

3. The mesenchymal stem cell separation device according to claim 2, wherein: Both the liquid injection pipe (311) and the liquid discharge pipe (312) are provided with a stop valve (34).

4. The mesenchymal stem cell separation device according to claim 3, wherein: A sealing rubber ring (14) is provided on the upper inner portion of the lower edge (11).

5. The mesenchymal stem cell separation device according to claim 4, wherein: An open groove (111) is provided at the lower portion of the lower edge (11).

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