In-vitro amplification kit for mesenchymal stem cells

Abstract

The utility model relates to an in-vitro amplification kit for mesenchymal stem cells, which comprises a kit body, reagent bottles and reagent tubes, the reagent bottles and the reagent tubes are arranged in the kit body, an in-vitro amplification culture box is further arranged in the kit body, and the in-vitro amplification culture box is integrally in a rectangular shape matched with the size of the kit body. The in-vitro amplification culture box is provided with a plurality of amplification wall fins perpendicular to the bottom face, the amplification wall fins are transversely arranged on the bottom face of the in-vitro amplification culture box in multiple rows, test solution diffusion channels are reserved between the amplification wall fins in each row, and liquid injection and extraction grooves are formed in the two sides of the in-vitro amplification culture box. The in-vitro amplification kit for the mesenchymal stem cells integrates an amplification culture box which adapts to the in-vitro amplification adherent growth characteristics of the mesenchymal stem cells and is convenient for reagent replacement, and a stepped complete in-vitro amplification operation platform is formed after the box body is unfolded, so that tissue cells are convenient to move and place, a test solution is convenient to inject and extract, and a centrifugal tube is convenient to take and place for observation; the functionality of the product is enriched, and convenience is provided for in-vitro amplification culture 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 an in vitro amplification kit for mesenchymal stem cells. Background Art

[0002] An in vitro amplification kit for mesenchymal stem cells is a reagent set box containing a cell culture solution and an induction amplification solution that are combined into a set. It contains various reagent solutions with fixed dosages required for in vitro culture and amplification of separated mesenchymal stem cell clusters, which are respectively placed in independent reagent bottles, and then these reagent bottles are combined into a set and uniformly placed in a box body to form a product. The introduced mesenchymal stem cell clusters complete the quantity expansion of mesenchymal stem cells through multiple centrifugation purifications and culture proliferations. The truly existing in vitro culture and amplification kits for mesenchymal stem cells on the market are generally ordinary carton packages that gather multiple reagent bottles of induction amplification solutions, with single functionality. Among the innovative cases of kits in the existing technical solutions, such as the innovative technologies of amplification kits like CN215939766U, CN211970177U, and CN209685807U, motor and other driving devices are added at the bottom or other positions of the kit to make the reagent bottles placed in the box (or case) rotate and vibrate, so that they have a mixing function. In addition to making the kit structure complex and increasing the cost, under the common knowledge in this field, reagent bottles are only used to hold reagents, and the purification and amplification of cells are not carried out in the reagent bottles but in centrifuge tubes and culture dishes respectively. Each reagent needs to be aspirated and transferred to the centrifuge tube or culture dish multiple times according to the steps when needed, and the inducer needs to be aspirated out (the inducer) and the culture solution needs to be updated regularly (the culture solution) after the steps are completed. Therefore, the above innovative functions actually do not have practicality. Content of the Utility Model

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

[0004] The in vitro amplification kit for mesenchymal stem cells includes a rectangular box body, and a reagent bottle a for accommodating serum culture medium, a reagent bottle b for containing a rinsing agent, and several reagent tubes for accommodating various growth factor solutions and antibacterial agents placed in the box body. An in vitro amplification culture box is also provided in the box body. Among them, the in vitro amplification culture box is integrally rectangular and adapted to the size of the box body. The in vitro amplification culture box is provided with several amplification wall fins perpendicular to the bottom surface. The amplification wall fins are arranged in multiple horizontal rows on the bottom surface of the in vitro amplification culture box, and a test solution diffusion channel is left between each row of amplification wall fins. Injection and extraction grooves are provided on both sides of the in vitro 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 in vitro 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] Further, 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 tapes 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] Further, 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] The in vitro amplification kit for mesenchymal stem cells of the present utility model overcomes the defect of the existing in vitro cell amplification kit with single functionality. In addition to having the function of reagent storage, it also integrates an amplification culture box that adapts to the adherent growth characteristics of mesenchymal stem cells in vitro and facilitates reagent replacement. After the box body is unfolded, it forms a stepped complete in vitro amplification operation platform. The front-placed culture box and the 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 being practical, the functionality of the product is enriched, providing convenience for the in vitro amplification culture practice operation of mesenchymal stem cells. Description of the Drawings

[0010] The following further describes the in vitro amplification kit for mesenchymal stem cells of the present utility model with reference to the drawings:

[0011] Figure 1 It is a three-dimensional structural schematic diagram of the in vitro amplification kit for mesenchymal stem cells of the present invention;

[0012] Figure 2 isFigure 1 Rear 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 Reference diagram of the first action of the usage state of the in vitro expansion kit for mesenchymal stem cells;

[0016] Figure 6 is Figure 5 Rear view;

[0017] Figure 7 is Reference diagram of the second action of the usage state of the in vitro expansion kit for mesenchymal stem cells.

[0018] In the figure:

[0019] 1 - Box body; 11 - Front 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 - In vitro amplification culture box; 51 - Amplification wall fins, 52 - Test solution diffusion channel, 53 - Injection and extraction tank;

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

[0025] In the present utility model, unless otherwise clearly specified and defined, 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 integrated; it can be directly connected, or indirectly connected through an intermediate medium, and 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", "rear", "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 in vitro amplification kit for mesenchymal stem cells of the present invention includes a rectangular box body 1, and a reagent bottle a 2 for containing serum culture medium, a reagent bottle b 3 for containing a flushing agent, and a number of reagent tubes 4 for containing various growth factor solutions and antibacterial agents placed in the box body 1. An in vitro amplification culture box 5 is also provided in the box body 1. Among them, the in vitro amplification culture box 5 is integrally rectangular and adapted to the size of the box body 1. The in vitro amplification culture box 5 is provided with a number of amplification wall fins 51 perpendicular to the bottom surface. The amplification wall fins 51 are arranged in multiple horizontal rows on the bottom surface of the in vitro amplification culture box 5. A test solution diffusion channel 52 is left between each row of amplification wall fins 51. Injection and extraction grooves 53 are provided on both sides of the in vitro amplification culture box 5. The separated and purified mesenchymal stem cell mass is dispersed and inoculated at the roots of each amplification wall fin. 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 liquid extraction of the pipette. For the convenience of 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. Repeating this several times can achieve uniform diffusion and soaking of the test solution on the plate on the tissue sample. Under the enhancement of growth factors and the protection of antibacterial agents, a large number of mesenchymal stem cells adhere and grow. The uniformly dispersed amplification wall fins provide a large amount of vertical wall area for cell adhesion and growth, improving the efficiency of cell in vitro proliferation.

[0029] Embodiment 2: In vitro amplification kit for mesenchymal stem cells. On both sides of the front panel 11 of the box body 1, there are virtual cutting lines 111. The bottom surface of the in vitro 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 pressing 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 pressing 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. Behind the outer top cover 12, there is a V-shaped support bar 123 for supporting the lower part of the outer top cover 12 in the unfolded state. On both sides of the V-shaped support bar 123, there is a double-sided coated 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 sized 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 cutting 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 an in vitro amplification 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 cutting line to fully expand the box body. The (centrifuge tube) tube placement structure, the reagent bottle / tube, and the culture box form a stepped structure, constituting an in vitro amplification operation platform with complete functions. Disperse and inoculate the separated and purified mesenchymal stem cell mass to the roots of each amplification wall fins. Use a pipette to draw each reagent according to the culture steps and inject it into one side of the injection and extraction groove, or inject liquid into both side grooves respectively and extract from the opposite side groove until the liquid level rises to soak the bottom surface of the culture box, realizing uniform diffusion and infiltration of the test solution on the plate to the inoculated cell mass; under the synergistic effect of growth factors and the protection of antibacterial agents, mesenchymal stem cells grow adherently in large numbers. The uniformly distributed amplification wall fins provide a large amount of vertical wall area for cell adherent growth and proliferation, improving the in vitro proliferation efficiency of cells. After amplification for a certain period of time, filter and add to a centrifuge tube, and centrifuge in a centrifuge to separate the cell mass from the culture solution containing metabolic wastes. Discard the upper layer of the old culture solution containing a large amount of metabolites. 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, and then disperse and inoculate again to the roots of each amplification wall fins, inject new culture solution and continue to culture until the required cell amount is amplified.

[0031] The utility model relates to an in vitro amplification kit for mesenchymal stem cells, which overcomes the defect of the existing cell in vitro amplification kit with single functionality. In addition to having the function of reagent storage, it also integrates an amplification culture box that adapts to the adherent growth characteristics of mesenchymal stem cells in vitro amplification and facilitates reagent replacement. After the box body is unfolded, it forms a stepped complete in vitro amplification operation platform. The front-placed culture box and the high-position centrifuge tube are convenient for tissue cell transfer and placement, reagent injection and extraction, and centrifuge tube removal and placement for observation. On the premise of having practicability, it enriches the functionality of the product and provides convenience for the in vitro amplification culture practice operation of mesenchymal stem cells.

[0032] The above description shows the main features, basic principles, and advantages of the utility model. For those skilled in the art, it is obvious that the utility model is not limited to the details of the above exemplary embodiments or examples, and without departing from the spirit or basic characteristics of the utility model, the utility model can be implemented in other specific forms. Therefore, the above embodiments or examples should be regarded as exemplary and non-restrictive. The scope of the utility model 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 utility model. Any reference signs in the claims should not be regarded as limiting the claims involved.

[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. An in vitro amplification kit for mesenchymal stem cells, comprising a rectangular box body (1), a reagent bottle a (2) placed in the box body (1) for containing serum culture medium, a reagent bottle b (3) for containing a rinsing agent, and a number of reagent tubes (4) placed in the box body (1) for containing various growth factor solutions and antibacterial agents, characterized in that: An in vitro amplification culture box (5) is further provided inside the box body (1), where the in vitro amplification culture box (5) is integrally rectangular and adapted to the size of the box body (1). The in vitro amplification culture box (5) is provided with a number of amplification wall fins (51) perpendicular to the bottom surface. The amplification wall fins (51) are arranged in multiple horizontal rows on the bottom surface of the in vitro amplification culture box (5). A test solution diffusion channel (52) is left between each row of amplification wall fins (51). Injection and extraction grooves (53) are provided on both sides of the in vitro amplification culture box (5).

2. The in vitro amplification kit for mesenchymal stem cells 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). The bottom surface of the in vitro 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 in vitro amplification kit for mesenchymal stem cells according to claim 2, characterized in that: 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) pressed outward is provided 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) pressed inward is provided 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 in vitro amplification kit for mesenchymal stem cells according to claim 3, characterized in that: A V-shaped support strip (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). Laminated double-sided adhesives (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 strip (123).

5. The in vitro amplification kit for mesenchymal stem cells according to claim 4, characterized in that: 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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