Mesenchymal stem cell separation and amplification kit

Abstract

The utility model relates to a mesenchymal stem cell separation and amplification kit which comprises a rectangular box body, a reagent bottle a, a reagent bottle b and a plurality of reagent tubes, the reagent bottle a, the reagent bottle b and the reagent tubes are arranged in the box body, the reagent bottle a is used for containing a culture solution, the reagent bottle b is used for containing a flushing agent, the reagent tubes are used for containing an inducer, and a separation and amplification culture box is further arranged in the box body. The whole separation and amplification culture box is in a rectangle shape matched with the size of a box body, a tissue access area is reserved in the center of the separation and amplification culture box, a plurality of amplification vertical plates extending from the center to the periphery in a radial mode are distributed on the periphery of the tissue access area in the circumferential direction, and the amplification vertical plates are of a trapezoidal structure gradually widened from inside to outside. Liquid injection and extraction grooves are formed in the two sides of the separation and amplification culture box. The mesenchymal stem cell separation and amplification kit integrates an amplification culture box adapting to the separation and amplification characteristics of mesenchymal stem cells, and a stepped complete separation and culture operation platform is formed after the box body is unfolded, so that the functionality of a product is enriched, and convenience is provided for separation and amplification practical operation of the mesenchymal stem cells.

Description

Technical Field

[0001] The utility model relates to the field of biotechnology and equipment, and particularly relates to a mesenchymal stem cell separation and amplification kit. Background Art

[0002] A mesenchymal stem cell separation and amplification kit is a reagent set box equipped with a combined induction and amplification solution. It contains various fixed-dose reagent solutions required for separating and amplifying mesenchymal stem cells from tissues, which are respectively placed in independent reagent bottles. Then these reagent bottles are combined into a set and uniformly placed in a box body to form a product. After the fragmented ex vivo tissue undergoes repeated centrifugation precipitation separation and adherent growth and proliferation, the expansion and purification of mesenchymal stem cells are completed. The truly existing mesenchymal stem cell separation and amplification kits on the market are generally ordinary cardboard packages that collect and store multiple reagent bottles of induction and amplification solutions, with single functionality. Among the innovative cases of kits in the existing technical solutions, such as the amplification kit innovative technologies of CN215939766U, CN211970177U, CN209685807U, etc., motor-driven devices such as motors are added at the bottom or other positions of the kit to rotate and shake the reagent bottles placed in the box (container), so that they have a mixing function. In addition to making the kit structure complex and increasing the cost, in the common knowledge of this field, reagent bottles are only used to hold reagents. The separation and amplification do not occur in the reagent bottles but in centrifuge tubes and culture dishes. The reagents need to be aspirated and transferred to centrifuge tubes and cultures step by step and then aspirated and replaced. Therefore, the above innovative functions actually do not have practicality. Summary of the Utility Model

[0003] The technical problem to be solved by the utility model is to overcome the defect of the single functionality of the existing cell separation and amplification kit, and provide a mesenchymal stem cell separation and amplification kit. In addition to having the function of holding reagents, it also integrates an amplification culture box that adapts to the characteristics of mesenchymal stem cell separation and amplification from the inside out and is convenient for reagent replacement. After the box body is unfolded, it forms a stepped complete separation and culture operation platform. The front-placed culture box and high-position centrifuge tube are convenient for tissue cell transfer and placement, test solution injection and extraction, and centrifuge tube removal and placement for observation. On the premise of having practicality, it enriches the functionality of the product and provides convenience for the practical operation of mesenchymal stem cell separation and amplification.

[0004] The mesenchymal stem cell isolation and amplification kit includes a rectangular box body, and a reagent bottle a for containing culture medium, a reagent bottle b for containing rinsing agent, and several reagent tubes for containing inducing agent placed in the box body. A separation and amplification culture box is also provided in the box body. Among them, the separation and amplification culture box is integrally rectangular and adapted to the size of the box body. A tissue access area is left in the central part of the separation and amplification culture box, and several amplification vertical plates extending radially from the center to the periphery are arranged circumferentially outside the tissue access area. The amplification vertical plates form a trapezoidal structure that gradually widens from the inside to the outside, and injection and extraction grooves are provided on both sides of the separation and amplification culture box.

[0005] Optimally, virtual cutting lines are provided on both sides of the front panel of the box body, and the bottom surface of the separation and amplification culture box at the position of the injection and extraction grooves is pasted on the inner surface of the front panel.

[0006] Optimally, the top cover of the box body includes an inner layer and an outer layer. The front and rear edges of the outer layer top cover and the inner layer top cover are connected as a whole. Among them, a crease is pressed outward in the center of the outer layer top cover, and a small hole is opened in the rear area of the outer layer top cover; a crease is pressed inward in the center of the inner layer top cover, and a large hole corresponding to the small hole is opened in the front area of the inner layer top cover.

[0007] Furthermore, a V-shaped support bar for supporting the lower part of the outer layer top cover in the unfolded state is provided behind the outer layer top cover, and double-sided coated films for pasting and connecting the back panel of the box body in the unfolded state are provided on both sides of the V-shaped support bar.

[0008] Furthermore, a sponge container seat for fixing the reagent bottle a, the reagent bottle b, and the reagent tubes is provided in the box body.

[0009] A mesenchymal stem cell isolation and amplification kit of the present utility model overcomes the defect of the existing cell isolation and amplification kit with a single function. In addition to the function of containing reagents, it also integrates an amplification culture box that adapts to the characteristics of mesenchymal stem cell separation and amplification from the inside to the outside and facilitates reagent replacement. After the box body is unfolded, it forms a stepped and complete separation and culture operation platform. The front-placed culture box and the high-position centrifuge tube are convenient for tissue cell transfer and placement, solution injection and extraction, and centrifuge tube removal and placement for observation. On the premise of being practical, it enriches the functionality of the product and provides convenience for the practical operation of mesenchymal stem cell separation and amplification. Description of the Drawings

[0010] The following further describes a mesenchymal stem cell isolation and amplification kit of the present utility model with reference to the drawings:

[0011] Figure 1 is a three-dimensional structural schematic diagram of the mesenchymal stem cell isolation and amplification kit;

[0012] Figure 2 is Figure 1Rear view;

[0013] Figure 3 is Figure 1 Schematic diagram of the three-dimensional structure in the unfolded state;

[0014] Figure 4 is Figure 3 Rear view;

[0015] Figure 5 is a reference diagram of the first action of the usage state of the mesenchymal stem cell isolation and amplification kit;

[0016] Figure 6 is Figure 5 Rear view;

[0017] Figure 7 is a reference diagram of the second action of the usage state of the mesenchymal stem cell isolation and amplification kit.

[0018] In the figure:

[0019] 1 - box body; 11 - front side panel, 12 - outer top cover, 13 - inner top cover; 111 - virtual knife line, 121 - outward crease, 122 - small hole, 123 - V-shaped support bar, 124 - coated double-sided tape, 131 - inward crease, 132 - large hole;

[0020] 2 - reagent bottle a;

[0021] 3 - reagent bottle b;

[0022] 4 - reagent tube;

[0023] 5 - isolation and amplification culture box; 51 - tissue access area, 52 - amplification upright plate, 53 - injection and extraction groove;

[0024] 6 - sponge container seat. Detailed implementation manners

[0025] In the present utility model, unless otherwise clearly defined and limited, the terms "installation", "connection", "connection", "fixation" and other terms should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral body; it can be directly connected, or indirectly connected through an intermediate medium, and it can be the internal communication of two components or the interaction relationship between two components. For those of ordinary skill in the art, the specific meanings of the above terms in the present utility model can be understood according to specific circumstances.

[0026] In the description of the present utility model, it should be understood that the orientation or positional relationship indicated by the terms "left", "right", "front", "back", "top", "bottom", "inner", "outer", etc. are all based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present utility model and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation to the present utility model.

[0027] The following uses specific embodiments to further describe the technical solution of the present utility model, but the protection scope of the present utility model is not limited to the following embodiments.

[0028] Embodiment 1: As Figures 1 to 4 shown, the present mesenchymal stem cell isolation and amplification kit includes a rectangular box body 1, and a reagent bottle a 2 for containing culture solution, a reagent bottle b 3 for containing a rinsing agent, and a number of reagent tubes 4 for containing an inducer, which are placed in the box body 1. A separation and amplification culture box 5 is also provided in the box body 1. Among them, the separation and amplification culture box 5 is integrally rectangular and adapted to the size of the box body 1. A tissue access area 51 is left in the central part of the separation and amplification culture box 5, and a number of amplification vertical plates 52 extending radially from the center to the periphery are arranged circumferentially outside the tissue access area 51. The amplification vertical plates 52 are trapezoidal structures that gradually widen from the inside to the outside. Injection and extraction grooves 53 are provided on both sides of the separation and amplification culture box 5. The crushed tissue sample is inoculated into the tissue access area. A pipette is used to extract each reagent according to the culture steps and inject it into one side of the injection and extraction groove until the liquid level rises to soak the tissue sample. When extracting, the reagent is extracted from the injection and extraction grooves on both sides. This method can prevent the tissue sample from being directly impacted by the hydraulic pressure of the liquid outlet and extraction of the pipette. To facilitate the uniform diffusion of the test solution, the liquid can be injected into the grooves on both sides respectively, and extracted from the groove on the opposite side. After repeating several times, the test solution on the plate can be uniformly diffused to soak the tissue sample. Under the induction of the test solution, a large number of mesenchymal stem cells adhere and grow, and spread outwards. The trapezoidally widened vertical plates provide a large amount of wall area for the cells to adhere and grow in clusters, improving the efficiency of cell separation and purification.

[0029] Embodiment 2: On both sides of the front panel 11 of the box body 1 of the mesenchymal stem cell isolation and amplification kit, there are virtual knife lines 111. The bottom surface of the isolation and amplification culture box 5 at the position of the injection and extraction groove 53 is pasted on the inner surface of the front panel 11. The top cover of the box body 1 includes an inner layer and an outer layer. The front and rear edges of the outer top cover 12 and the inner top cover 13 are connected as a whole. Among them, a crease 121 that presses outward is formed in the center of the outer top cover 12, and a small hole 122 is opened in the rear area of the outer top cover 12; a crease 131 that presses inward is formed in the center of the inner top cover 13, and a large hole 132 corresponding to the small hole 122 is opened in the front area of the inner top cover 13. A V-shaped support bar 123 for supporting the lower part of the outer top cover 12 in the unfolded state is provided behind the outer top cover 12. On both sides of the V-shaped support bar 123, there is a double-sided film adhesive 124 for pasting and connecting to the back panel of the box body 1 in the unfolded state. After opening the lid, the inner and outer top covers are folded in opposite directions to form a frame, and the corresponding large and small holes on the upper and lower surfaces of the frame form a tube placement structure for placing centrifuge tubes. Inside the box body 1, there is a sponge container seat 6 for fixing the reagent bottle a2, the reagent bottle b3, and the reagent tube 4. The sponge container seat is pasted inside the box body, and slot holes adapted to each reagent bottle / tube are opened thereon. After opening the lid and tearing the virtual knife line to expand the box body, the (centrifuge tube) tube placement structure, the reagent bottle / tube, and the culture box form a stepped structure, constituting a separation and culture operation platform with complete functions. The remaining structures and components are as described in Embodiment 1 and will not be repeated.

[0030] During use: As Figure 5 , 6 shown, open and expand the inner and outer top covers, and paste both sides of the V-shaped support bar on the back panel of the box body; as Figure 7 shown, then tear the virtual knife line to fully expand the box body, and the (centrifuge tube) tube placement structure, the reagent bottle / tube, and the culture box form a stepped structure, constituting a separation and culture operation platform with complete functions. The fragmented tissue sample is inoculated into the tissue access area of the culture box, and each reagent is extracted with a pipette according to the culture steps and injected into one side of the injection and extraction groove, or liquid is injected into both side grooves respectively, and extracted from the opposite side groove until the liquid level rises to soak the tissue sample; under the induction of the test solution, a large number of mesenchymal stem cells adhere and grow, and spread outward. The trapezoidally widened vertical plate provides a large amount of vertical wall area for the cell clusters to adhere and grow, improving the efficiency of cell separation and purification; after the cell clusters are amplified to a certain number, filter and add them to a centrifuge tube and separate them in a centrifuge. After discarding the supernatant, place the pipette into the tube placement structure formed by the large and small holes of the inner and outer top covers, add a rinsing agent to rinse, soak with a culture solution, and then pour it back into the tissue access area of the culture box to continue culturing until the purified cells are amplified to the required cell quantity.

[0031] The Kit for Isolating and Amplifying Bone Marrow Mesenchymal Stem Cells overcomes the defect of the existing cell isolation and amplification kits with single functionality. In addition to the function of containing reagents, it integrates an amplification culture box that adapts to the characteristics of the isolation and amplification of mesenchymal stem cells from the inside out and facilitates the replacement of reagents. After the box body is unfolded, it forms a stepped and complete separation and culture operation platform. The front-placed culture box and the high-position centrifuge tube are convenient for transferring and placing tissue cells, injecting and extracting test solutions, and taking and placing the centrifuge tube for observation. On the premise of being practical, it enriches the functionality of the product and provides convenience for the practical operation of isolating and amplifying mesenchymal stem cells.

[0032] The above description shows the main features, basic principles, and advantages of the present invention. For those skilled in the art, it is obvious that the present invention is not limited to the details of the above exemplary embodiments or examples, and without departing from the spirit or basic characteristics of the present invention, the present invention can be implemented in other specific forms. Therefore, the above embodiments or examples should be regarded as exemplary and non-limiting. The scope of the present invention is defined by the appended claims rather than the above description. Therefore, all changes falling within the meaning and scope of the equivalent elements of the claims are intended to be included in the present invention. Any reference signs in the claims should not be regarded as limiting the claimed invention.

[0033] In addition, it should be understood that although this specification is described according to embodiments, not every embodiment only contains an independent technical solution. This narrative way of the specification is only for clarity. Those skilled in the art should regard the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A mesenchymal stem cell isolation and amplification kit, comprising a rectangular box body (1), a reagent bottle a (2) for containing a culture solution, a reagent bottle b (3) for containing a rinsing agent, and a number of reagent tubes (4) for containing an inducer, all of which are placed in the box body (1), and characterized in that: Inside the box body (1), there is also a separation amplification culture box (5), where The separation amplification culture box (5) is generally rectangular in shape and adapted to the size of the box body (1). There is a tissue access area (51) left in the central part of the separation amplification culture box (5), and several amplification vertical plates (52) radially extending from the center to the surrounding are arranged circumferentially around the tissue access area (51). The amplification vertical plates (52) are trapezoidally structured to be gradually wider from the inside to the outside, and injection and extraction grooves (53) are provided on both sides of the separation amplification culture box (5).

2. The mesenchymal stem cell isolation and amplification kit according to claim 1, characterized in that: Virtual cutting lines (111) are provided on both sides of the front side panel (11) of the box body (1), and the bottom surface of the separation amplification culture box (5) at the position of the injection and extraction grooves (53) is pasted on the inner surface of the front side panel (11).

3. The mesenchymal stem cell isolation and amplification kit according to claim 2, wherein: The top cover of the box body (1) includes an inner layer and an outer layer. The front and rear edges of the outer layer top cover (12) and the inner layer top cover (13) are connected as a whole. Among them, a crease (121) pressing outward is formed in the center of the outer layer top cover (12), and a small hole (122) is opened in the rear area of the outer layer top cover (12); a crease (131) pressing inward is formed in the center of the inner layer top cover (13), and a large hole (132) corresponding to the small hole (122) is opened in the front area of the inner layer top cover (13).

4. The mesenchymal stem cell isolation and amplification kit according to claim 3, wherein: A V-shaped support bar (123) for supporting the lower part of the outer layer top cover (12) in the unfolded state is provided behind the outer layer top cover (12), and double-sided adhesive films (124) for pasting and connecting the back side panel of the box body (1) in the unfolded state are provided on both sides of the V-shaped support bar (123).

5. The mesenchymal stem cell isolation and amplification kit according to claim 4, wherein: A sponge container seat (6) for fixing the reagent bottle a (2), the reagent bottle b (3), and the reagent tube (4) is provided inside the box body (1).

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