Heart valve type organ cultivation device and use method thereof

Abstract

The invention relates to the technical field of cell culture, in particular to a heart valve type organ culture device and a use method thereof. Which comprises a cultivation box body and a sealing cover, the cultivation box body is fixed, the sealing cover is arranged on one side of the cultivation box body, and the device is characterized by further comprising a fixing device, a seed cell culture device and a cell planting device, the fixing device is used for fixing the cell planting device and the seed cell culture device, the cell culture device is mounted on the left side of the culture box, and the cell culture device is used for reproduction of seed cells in an in-vitro environment until enough quantity is achieved for planting; the cell planting device is used for placing the cells on the stent for cultivation after the seed cells are fixed, so that the cells form a continuous cell layer on the surface of the stent until the culture of the heart valve is completed.

Description

Technical Field

[0001] The present invention relates to the technical field of cell cultivation, and in particular to a heart valve organoid cultivation device and a method for using the same. Background Art

[0002] The cultivation of artificial heart valves is of great significance to patients with heart disease. It not only provides new treatment options for patients who cannot undergo traditional surgical treatment due to valve disease, but also reduces surgical risks and postoperative complications. Valves cultivated through bioengineering can better simulate the functions of natural valves in the human body, improve the quality of life of patients, and may reduce the need for long-term anticoagulation therapy.

[0003] In the process of cultivating existing heart valves, it is usually necessary to first cultivate seed cells in an instrument, and then manually add substances such as nutrient solution. After the seed cell cultivation is completed, it is transferred to another instrument to plant the expanded seed cells on the decellularized stent, and then manually add nutrient solution to complete the cultivation of the heart valve. The above-mentioned method of manually adding nutrient solution will reduce work efficiency and increase the risk of culture medium infection, which will lead to cultivation failure.

[0004] Therefore, the present invention provides a heart valve organoid cultivation device and a method of using the same to solve the above-mentioned problems. Summary of the invention

[0005] Technical problem to be solved by the present invention: In the process of cultivating existing heart valves, it is usually necessary to first cultivate seed cells in an instrument, and then artificially add substances such as nutrient solution. After the seed cell cultivation is completed, it is transferred to another instrument to plant the amplified seed cells on the decellularized stent, and then artificially add nutrient solution to complete the cultivation of the heart valve. The above-mentioned method of artificially adding nutrient solution will reduce work efficiency and increase the risk of culture medium infection, which will lead to cultivation failure.

[0006] The present invention provides the following technical solutions: a heart valve organoid cultivation device, comprising a cultivation box body and a sealing cover, wherein the cultivation box body is fixed, and a sealing cover is provided on one side of the cultivation box body, and is characterized in that it also comprises a fixing device, a seed cell cultivation device and a cell implantation device, wherein the fixing device is installed inside the cultivation box body, the fixing device is used to fix the cell implantation device and the seed cell cultivation device, the cell cultivation device is installed on the left side of the cultivation box, and the cell cultivation device is used to reproduce the seed cells in an in vitro environment until a sufficient number is reached for planting; the cell implantation device is used to place the cells on a stent for cultivation after the seed cells are fixed, so that the cells form a continuous cell layer on the surface of the stent until the cultivation of the heart valve is completed.

[0007] Preferably, the fixing device comprises a fixing groove, a telescopic rod and a clamping block. The incubator body is provided with a fixing groove, two telescopic rods are arranged in the fixing groove, and the clamping block is arranged on the telescopic rod.

[0008] Preferably, the seed cell culture device comprises a No. 1 culture chamber, a No. 1 infusion tube and a No. 1 constant temperature heater, the No. 1 culture chamber is fixed in the incubator body by a fixing device, the No. 1 infusion tube is installed on one side of the No. 1 culture chamber, and a constant temperature heater is installed at the bottom of the No. 1 culture chamber.

[0009] Preferably, the first culture chamber is provided with an oxygen supply hole and a heating hole in an array outside, and the oxygen supply hole and the heating hole are connected to a glass cover outside; the oxygen supply hole and the heating hole are arranged in a cross array, and the oxygen supply hole and the heating hole are provided with a recovery hole outside. A solenoid valve is provided inside the recovery hole.

[0010] Preferably, the cell implantation device comprises a No. 2 culture chamber, a No. 2 infusion tube, a No. 3 infusion tube, a No. 2 constant temperature heater and a supporting device; the No. 2 culture chamber is installed in the incubator body; the No. 2 culture chamber is divided into two parts, an upper part and an lower part; and the No. 2 culture chamber is symmetrically installed with a supporting device; the No. 2 infusion tube and the No. 3 infusion tube are installed on both sides of the No. 2 culture chamber; and the No. 2 constant temperature heater is installed at the bottom of the No. 2 culture chamber.

[0011] Preferably, the supporting device comprises an adjustable column and a supporting gasket, the adjustable column is symmetrically installed on both sides of the No. 2 culture chamber, and an umbrella-shaped supporting gasket is installed on the adjustable column.

[0012] Preferably, a pull-out plate is provided in the incubator body, the fixing device, the seed cell culture device and the cell planting device are all installed on the pull-out plate, and the pull-out plate is provided with a handle.

[0013] A method for using a heart valve organoid cultivation device, the specific steps are as follows: S1: Open the sealing cover, place the No. 1 culture chamber in the fixing groove of the fixing device, and squeeze the telescopic rod in the fixing device backward until the No. 1 culture chamber is completely placed in and then rebounds and clamps; S2: After the No. 1 culture chamber is clamped, the hose for conveying nutrient solution in the incubator is inserted into the No. 1 infusion tube to fill a certain amount of nutrient solution, and then the No. 1 constant temperature heater is turned on to heat the No. 1 culture chamber. After heating to the specified temperature, the staff puts the seed cells into the No. 1 culture chamber, then buckles the glass cover and controls the corresponding oxygen supply machine to supply oxygen and heat the gas through the control device, thereby ensuring that the oxygen content and temperature in the No. 1 culture chamber are constant and starting to culture the seed cells in vitro; S3: After the seed cell culture is completed, the staff inserts the stent into the support gasket of the support device, and inserts the hose for delivering the nutrient solution in the incubator into the No. 2 infusion tube to first fill a certain amount of nutrient solution, and at the same time delivers the cross-linking agent through the No. 3 infusion tube; then the No. 2 constant temperature heater is turned on to heat the No. 2 culture chamber. After heating to the specified temperature, the staff puts the seed cells into the No. 2 culture chamber, and implants the amplified seed cells onto the decellularized stent, and forms a continuous cell layer on the surface of the stent through in vitro culture; S4: Once the cell layer is formed and cross-linked, the heart valve can be removed for further testing.

[0014] The beneficial effects of the present invention are as follows: 1. The present invention adopts a new seed cell culture device, which can automatically control the internal temperature during the cell culture process. At the same time, due to the setting of the No. 1 infusion tube, the nutrient solution can be automatically added to the No. 1 culture chamber, thereby improving the culture efficiency of the seed cells; and a glass cover is provided in the No. 1 culture chamber, and oxygenation holes and heating holes are provided in the glass cover, thereby ensuring sufficient oxygen during the culture process and stable temperature of the entire external environment.

[0015] 2. The present invention adopts a new cell implantation device, and a support device is arranged inside the No. 2 culture chamber. The support device is used to adaptively support the stent, thereby achieving its fixation. Then, nutrient solution and other required substances are automatically added to it through the No. 2 infusion tube and the No. 3 infusion tube. Then, the contact position between the stent and the seed cells is adjusted by moving the adjustable column, and then the amplified seed cells are implanted on the decellularized stent. Through in vitro culture, the cells form a continuous cell layer on the surface of the stent to complete the production of the heart valve. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0017] Figure 1 It is an overall schematic diagram of the present invention; Figure 2 It is a schematic diagram of the fixing device of the present invention; Figure 3 It is a schematic diagram of the seed cell culture device of the present invention; Figure 4 This is a schematic diagram of the recovery hole of the present invention; Figure 5 This is a schematic diagram of the interior of the cell implantation device of the present invention; Figure 6 It is a schematic diagram of the cell planting device of the present invention; Figure 7 It is a schematic diagram of a pull-out plate of the present invention; Figure 8 The figure is a flow chart of the method of the present invention.

[0018] In the figure: 1. incubator body; 11. pull-out plate; 12. handle; 2. sealing cover; 3. fixing device; 31. fixing slot; 32. telescopic rod; 33. clamping block; 4. seed cell culture device; 41. culture chamber No. 1; 42. infusion tube No. 1; 43. constant temperature heater No. 1; 44. oxygen supply hole; 45. heating hole; 46. recovery hole; 47. solenoid valve; 5. cell planting device; 51. culture chamber No. 2; 52. infusion tube No. 2; 53. infusion tube No. 3; 54. constant temperature heater No. 2; 55. supporting device; 551. adjustable column; 552. supporting gasket. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Therefore, the following detailed description of the embodiments of the present invention is not intended to limit the scope of the claimed invention, but merely represents some embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0020] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0021] In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", "back" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, or the positions or positional relationships in which the product of the invention is usually placed when in use. Such terms are only used to facilitate the description of the present invention and simplify the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as limiting the present invention.

[0022] It should also be noted that in the description of the present invention, unless otherwise clearly specified and limited, the terms "set", "install", "connect", and "connect" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0023] The disclosed embodiments are intended to solve the problem that in the existing heart valve cultivation process, seed cells usually need to be cultivated in an instrument first, and then nutrient solution and other substances are added manually. After the seed cell cultivation is completed, the seed cells are transferred to another instrument to plant the expanded seed cells on the decellularized stent, and then nutrient solution is added manually to complete the heart valve cultivation. The above-mentioned method of artificially adding nutrient solution will reduce work efficiency and increase the risk of culture medium infection, which will lead to the problem of cultivation failure. In view of this, the disclosed embodiment proposes a heart valve organoid cultivation device and a method of using the same, which adopts a new seed cell culture device, and can automatically control the internal temperature thereof during the cell culture process. At the same time, due to the setting of the No. 1 infusion tube, the nutrient solution can be automatically added to the No. 1 culture chamber, thereby improving the culture efficiency of the seed cells; and a glass cover is provided in the No. 1 culture chamber, and oxygenation holes and heating holes are provided in the glass cover, thereby ensuring sufficient oxygen and stable temperature of the entire external environment during the culture process; at the same time, a support device is provided inside the No. 2 culture chamber, and the support device is used to adaptively support the stent, thereby achieving its fixation, and then through the No. 2 infusion tube and the No. 3 infusion tube, nutrient solution and other required substances are automatically added to it, and then the contact position between the stent and the seed cells is adjusted by moving the adjustable column, and then the amplified seed cells are implanted on the decellularized stent, and the cells are cultured in vitro to form a continuous cell layer on the surface of the stent to complete the production of the heart valve.

[0024] like Figures 1 to 8As shown, a heart valve organoid cultivation device comprises a cultivation box body 1 and a sealing cover 2, wherein the cultivation box body 1 is fixed, and a sealing cover 2 is provided on one side of the cultivation box body 1, and is characterized in that it also comprises a fixing device 3, a seed cell cultivation device 4 and a cell planting device 5, wherein the fixing device 3 is installed inside the cultivation box body 1, and the fixing device 3 is used to fix the cell planting device 5 and the seed cell cultivation device 4, and the cell cultivation device is installed on the left side of the cultivation box, and the cell cultivation device is used to reproduce the seed cells in an in vitro environment until a sufficient number is reached for planting; the cell planting device 5 is used to place the cells on a stent for cultivation after the seed cells are fixed, so that the cells form a continuous cell layer on the surface of the stent until the cultivation of the heart valve is completed; By adopting the new seed cell culture device 4, the temperature inside it can be automatically controlled during the cell culture process. At the same time, due to the setting of the No. 1 infusion tube 42, the nutrient solution can be automatically added to the No. 1 culture chamber 41, thereby improving the culture efficiency of the seed cells; and a glass cover is provided in the No. 1 culture chamber 41, and an oxygenation hole and a heating hole 45 are provided in the glass cover, thereby ensuring sufficient oxygen and stable temperature of the entire external environment during the culture process; at the same time, a support device 55 is provided in the No. 2 culture chamber 51, and the support device 55 is used to adaptively support the stent to achieve its fixation, and then the nutrient solution and other required substances are automatically added to it through the No. 2 infusion tube 52 and the No. 3 infusion tube 53, and then the contact position between the stent and the seed cells is adjusted by moving the adjustable column 551, and then the amplified seed cells are implanted on the decellularized stent, and the cells are cultured in vitro to form a continuous cell layer on the surface of the stent to complete the production of the heart valve.

[0025] like Figure 2 As shown, the fixing device 3 includes a fixing groove 31, a telescopic rod 32 and a clamping block 33. The incubator body 1 is provided with a fixing groove 31, and the fixing groove 31 is used to place the No. 1 culture cavity 41 and the No. 2 culture cavity 51; two telescopic rods 32 are arranged in the fixing groove 31, and the telescopic rods 32 are used to telescope and cooperate with the clamping block 33 to clamp the No. 1 culture cavity 41 and the No. 2 culture cavity 51; the telescopic rod 32 is used to cooperate with the clamping block 33 to telescope after the clamping block 33 is squeezed; the clamping block 33 is arranged on the telescopic rod 32, and the clamping block 33 is used to clamp the No. 1 culture cavity 41 and the No. 2 culture cavity 51; During operation, the staff presses the No. 1 culture chamber 41 or the No. 2 culture chamber 51, so that the No. 1 culture chamber 41 or the No. 2 culture chamber 51 squeezes the clamping block 33. After the clamping block 33 is squeezed, it pushes the telescopic rod 32 to contract, so that the No. 1 culture chamber 41 or the No. 2 culture chamber 51 is clamped between the clamping blocks 33 to complete the fixing work; the above-mentioned fixing device 3 can be used to quickly fix the No. 1 culture chamber 41 or the No. 2 culture chamber 51, thereby improving work efficiency.

[0026] like Figure 3 to Figure 4 As shown, the seed cell culture device 4 includes a No. 1 culture chamber 41, a No. 1 infusion tube 42 and a No. 1 constant temperature heater 43. The No. 1 culture chamber 41 is fixed in the incubator body 1 by a fixing device 3, and the No. 1 culture chamber 41 is used to culture seed cells; the No. 1 infusion tube 42 is installed on one side of the No. 1 culture chamber 41, and the No. 1 infusion tube 42 is used to transport the nutrient solution required for seed cell expansion to the No. 1 culture chamber 41; a constant temperature heater is installed at the bottom of the No. 1 culture chamber 41, and the constant temperature heater is used to adjust the temperature in the No. 1 culture chamber 41, so that the seeds can be cultured better; During operation, the sealing cover 2 is opened, and the No. 1 culture chamber 41 is placed in the fixing groove 31 of the fixing device 3. The telescopic rod 32 in the fixing device 3 is squeezed backwards and rebounds and clamped after the No. 1 culture chamber 41 is completely placed therein; after the No. 1 culture chamber 41 is clamped, the hose for conveying the nutrient solution in the incubator is inserted into the No. 1 infusion tube 42 to first fill a certain amount of nutrient solution, and then the No. 1 constant temperature heater 43 is turned on to heat the No. 1 culture chamber 41. After heating to the specified temperature, the staff puts the seed cells into the No. 1 culture chamber 41, then buckles the glass cover and simultaneously operates the corresponding oxygen supply machine through the control device to supply oxygen and heat through gas, thereby ensuring that the oxygen content and temperature in the No. 1 culture chamber 41 are constant and starting to culture the seed cells in vitro; By adopting the new seed cell culture device 4, the internal temperature can be automatically controlled during the cell culture process. At the same time, due to the setting of the No. 1 infusion tube 42, the nutrient solution can be automatically added to the No. 1 culture chamber 41, thereby improving the culture efficiency of the seed cells; and a glass cover is provided in the No. 1 culture chamber 41, and oxygenation holes and heating holes 45 are provided in the glass cover, thereby ensuring sufficient oxygen during the culture process and stable temperature of the entire external environment.

[0027] like Figure 4As shown, the outer array of the No. 1 culture chamber 41 is provided with an oxygen supply hole 44 and a heating hole 45, the oxygen supply hole 44 is used to provide the oxygen required for seed culture; the heating hole 45 is used to heat the oxygen, thereby achieving uniform temperature control of the No. 1 culture chamber 41; the oxygen supply hole 44 and the heating hole 45 are connected to the outside with a glass cover; the glass cover is used to make the oxygen flow within the moving range, thereby achieving uniform heating; the oxygen supply holes 44 and the heating holes 45 are arranged in a cross array, and the cross array arrangement is used to enable the heating holes 45 to better dissipate heat from the oxygen supply holes 44; the oxygen supply holes 44 and the heating holes 45 are provided with recovery holes 46 on the outside; the recovery hole 46 is provided with a solenoid valve 47, and the solenoid valve 47 is used to open or close, thereby controlling the gas content in the glass cover at any time; like Figure 5 to Figure 6 As shown, the cell planting device 5 includes a No. 2 culture chamber 51, a No. 2 infusion tube 52, a No. 3 infusion tube 53, a No. 2 constant temperature heater 54 and a supporting device 55. The No. 2 culture chamber 51 is installed in the incubator body 1, and the No. 2 culture chamber 51 is used to cultivate seed cells on the support; the No. 2 culture chamber 51 is divided into two parts, an upper part and an lower part, and the No. 2 culture chamber 51 is symmetrically installed with a supporting device 55; the supporting device 55 is used to support the support to be planted, so that the seed cells can form a continuous cell layer on the surface of the support; the No. 2 infusion tube 52 and the No. 3 infusion tube 53 are installed on both sides of the No. 2 culture chamber 51; the No. 2 infusion tube 52 and the No. 3 infusion tube 53 are used to transport the nutrient solution to the inside of the No. 2 culture chamber 51; the No. 2 constant temperature heater 54 is installed at the bottom of the No. 2 culture chamber 51, and the No. 2 constant temperature heater 54 is used to heat the No. 2 culture chamber 51; During operation, the staff inserts the stent into the support pad 552 of the support device 55, and inserts the hose for conveying nutrient solution in the incubator into the No. 2 infusion tube 52 to first fill a certain amount of nutrient solution, and at the same time conveys the cross-linking agent through the No. 3 infusion tube 53; then the No. 2 constant temperature heater 54 is turned on to heat the No. 2 culture chamber 51. After heating to the specified temperature, the staff puts the seed cells into the No. 2 culture chamber 51, and implants the amplified seed cells onto the decellularized stent, and forms a continuous cell layer on the surface of the stent through in vitro culture. A supporting device 55 is provided inside the No. 2 culture chamber 51, and the stent is adaptively supported by the supporting device 55 to achieve fixation, and then nutrient solution and other required substances are automatically added to it through the No. 2 infusion tube 52 and the No. 3 infusion tube 53, and then the contact position between the stent and the seed cells is adjusted by moving the adjustable column 551, and then the amplified seed cells are implanted on the decellularized stent, and the cells are cultured in vitro to form a continuous cell layer on the surface of the stent to complete the production of the heart valve.

[0028] like Figure 5 As shown, the supporting device 55 includes an adjustable column 551 and a supporting gasket 552. The adjustable column 551 is symmetrically installed on both sides of the No. 2 culture chamber 51. The adjustable column 551 is installed with an umbrella-shaped supporting gasket 552. The adjustable column 551 is used to adjust the height between it and the culture solution according to different brackets; the umbrella-shaped supporting gasket 552 is used to support brackets of different diameters, thereby ensuring the stability of the bracket.

[0029] During operation, the staff inserts the umbrella-shaped support pads 552 into both ends of the support, and adjusts the height thereof through the adjustable column 551 so that it can be fully integrated with the nutrient solution, thereby allowing the seed cells to form a continuous cell layer on the surface of the support.

[0030] like Figure 7 As shown, a pull-out plate 11 is provided in the incubator body 1, and the pull-out plate 11 is used to pull out the cell planting device 5 and the seed cell culture device 4 from the incubator body 1, so as to facilitate the staff to pull out or put in the No. 1 culture chamber 41 and the No. 2 culture chamber 51; the fixing device 3, the seed cell culture device 4 and the cell planting device 5 are all installed on the pull-out plate 11, and the pull-out plate 11 is provided with a handle 12, and the handle 12 is used to facilitate the staff to pull out the pull-out plate 11.

[0031] like Figure 8 As shown, S1: open the sealing cover 2, place the No. 1 culture chamber 41 in the fixing groove 31 of the fixing device 3, and the telescopic rod 32 in the fixing device 3 is squeezed backwards and rebounds and clamped after the No. 1 culture chamber 41 is completely placed in; S2: After the No. 1 culture chamber 41 is clamped, the hose for conveying the nutrient solution in the incubator is inserted into the No. 1 infusion tube 42 to fill a certain amount of nutrient solution, and then the No. 1 constant temperature heater 43 is turned on to heat the No. 1 culture chamber 41. After heating to the specified temperature, the staff puts the seed cells into the No. 1 culture chamber 41, then buckles the glass cover and controls the corresponding oxygen supply machine to supply oxygen and heat the gas through gas, thereby ensuring that the oxygen content and temperature in the No. 1 culture chamber 41 are constant and starting to culture the seed cells in vitro; S3: After the seed cell culture is completed, the staff inserts the stent into the support pad 552 of the support device 55, and inserts the hose for conveying the nutrient solution in the incubator into the No. 2 infusion tube 52 to first fill a certain amount of nutrient solution, and at the same time conveys the cross-linking agent through the No. 3 infusion tube 53; then the No. 2 constant temperature heater 54 is turned on to heat the No. 2 culture chamber 51. After heating to the specified temperature, the staff puts the seed cells into the No. 2 culture chamber 51, and implants the amplified seed cells onto the decellularized stent, and forms a continuous cell layer on the surface of the stent through in vitro culture; S4: Once the cell layer is formed and cross-linked, the heart valve can be removed for further testing.

[0032] The overall working process is: open the sealing cover 2, place the No. 1 culture chamber 41 in the fixing groove 31 of the fixing device 3, and the telescopic rod 32 in the fixing device 3 is squeezed backwards and rebounded and clamped after the No. 1 culture chamber 41 is completely placed in; after the No. 1 culture chamber 41 is clamped, the hose for conveying nutrient solution in the incubator is inserted into the No. 1 infusion tube 42 to first fill a certain amount of nutrient solution, and then the No. 1 constant temperature heater 43 is turned on to heat the No. 1 culture chamber 41. After heating to the specified temperature, the staff puts the seed cells into the No. 1 culture chamber 41, and then buckles the glass cover and simultaneously operates the corresponding oxygen supply machine through the control device to supply oxygen and heat through gas, thereby ensuring that the oxygen content and temperature in the No. 1 culture chamber 41 are constant. The seed cells are cultured in vitro; after the seed cells are cultured, the staff inserts the stent into the support pad 552 of the support device 55, and inserts the hose for conveying the nutrient solution in the incubator into the No. 2 infusion tube 52 to first fill a certain amount of nutrient solution, and at the same time conveys the cross-linking agent through the No. 3 infusion tube 53; then the No. 2 constant temperature heater 54 is turned on to heat the No. 2 culture chamber 51. After heating to the specified temperature, the staff puts the seed cells into the No. 2 culture chamber 51, and implants the amplified seed cells onto the decellularized stent, and forms a continuous cell layer on the surface of the stent through in vitro culture; after the cell layer is formed and cross-linking is completed, the heart valve can be taken out for further testing.

[0033] Those skilled in the art should understand that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, and these changes and improvements fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A heart valve organoid cultivation device, comprising a cultivation box body (1) and a sealing cover (2), wherein the cultivation box body (1) is fixed, and a sealing cover (2) is provided on one side of the cultivation box body (1), characterized in that It also comprises a fixing device (3), a seed cell culture device (4) and a cell planting device (5), wherein the fixing device (3) is installed inside the incubator body (1), the fixing device (3) is used to fix the cell planting device (5) and the seed cell culture device (4), the cell culture device is installed on the left side of the incubator, and the cell culture device is used to reproduce the seed cells in an in vitro environment until a sufficient number is reached for planting; the cell planting device (5) is used to place the cells on a stent for cultivation after the seed cells are fixed, so that the cells form a continuous cell layer on the surface of the stent until the cultivation of the heart valve is completed.

2. A heart valve organoid cultivation device according to claim 1, characterized in that: The fixing device (3) comprises a fixing groove (31), a telescopic rod (32) and a clamping block (33); the incubator body (1) is provided with a fixing groove (31), two telescopic rods (32) are arranged in the fixing groove (31), and the clamping block (33) is arranged on the telescopic rod (32).

3. A heart valve organoid cultivation device according to claim 2, characterized in that: The seed cell culture device (4) comprises a No. 1 culture chamber (41), a No. 1 infusion tube (42) and a No. 1 constant temperature heater (43); the No. 1 culture chamber (41) is fixed in the incubator body (1) via a fixing device (3); the No. 1 infusion tube (42) is installed on one side of the No. 1 culture chamber (41); and a constant temperature heater is installed at the bottom of the No. 1 culture chamber (41).

4. A heart valve organoid cultivation device according to claim 3, characterized in that: The first culture chamber (41) is provided with an oxygen supply hole (44) and a heating hole (45) in an array outside, and the oxygen supply hole (44) and the heating hole (45) are connected to a glass cover outside; the oxygen supply hole (44) and the heating hole (45) are arranged in a cross array, and the oxygen supply hole (44) and the heating hole (45) are provided with a recovery hole (46) outside. A solenoid valve (47) is provided inside the recovery hole (46).

5. A heart valve organoid cultivation device according to claim 4, characterized in that: The cell implantation device (5) comprises a No. 2 culture chamber (51), a No. 2 infusion tube (52), a No. 3 infusion tube (53), a No. 2 constant temperature heater (54) and a support device (55); the No. 2 culture chamber (51) is installed in the incubator body (1); the No. 2 culture chamber (51) is divided into an upper and a lower part; and the No. 2 culture chamber (51) is symmetrically installed with the support device (55); the No. 2 infusion tube (52) and the No. 3 infusion tube (53) are installed on both sides of the No. 2 culture chamber (51); and the No. 2 constant temperature heater (54) is installed at the bottom of the No. 2 culture chamber (51).

6. A heart valve organoid cultivation device according to claim 5, characterized in that: The supporting device (55) comprises an adjustable column (551) and a supporting gasket (552); the adjustable column (551) is symmetrically mounted on both sides of the No. 2 culture chamber (51); and an umbrella-shaped supporting gasket (552) is mounted on the adjustable column (551).

7. A heart valve organoid cultivation device according to claim 6, characterized in that: A pull-out plate (11) is provided in the incubator body (1); the fixing device (3), the seed cell culture device (4) and the cell planting device (5) are all mounted on the pull-out plate (11); and a handle (12) is provided on the pull-out plate (11).

8. A method for using a heart valve organoid cultivation device, characterized in that: Using a heart valve organoid cultivation device as claimed in any one of claims 1 to 7, the specific steps are as follows: S1: Open the sealing cover (2), place the No. 1 culture chamber (41) in the fixing groove (31) of the fixing device (3), and squeeze the telescopic rod (32) in the fixing device (3) backwards until the No. 1 culture chamber (41) is completely placed in the culture chamber and then rebounds and clamps; S2: After the No. 1 culture chamber (41) is clamped, the hose for conveying the nutrient solution in the incubator is inserted into the No. 1 infusion tube (42) to first fill a certain amount of nutrient solution, and then the No. 1 constant temperature heater (43) is turned on to heat the No. 1 culture chamber (41). After the chamber is heated to a specified temperature, the staff puts the seed cells into the No. 1 culture chamber (41), then puts on the glass cover and simultaneously operates the corresponding oxygen supply machine through the control device to supply oxygen and heat the chamber through gas, thereby ensuring that the oxygen content and temperature in the No. 1 culture chamber (41) are constant and starting to culture the seed cells in vitro; S3: After the seed cell culture is completed, the staff inserts the scaffold into the support pad (552) of the support device (55), and inserts the hose for conveying the nutrient solution in the incubator into the No. 2 infusion tube (52) to first fill a certain amount of nutrient solution, and at the same time conveys the cross-linking agent through the No. 3 infusion tube (53); then the No. 2 constant temperature heater (54) is turned on to heat the No. 2 culture chamber (51). After heating to a specified temperature, the staff puts the seed cells into the No. 2 culture chamber (51), and implants the amplified seed cells onto the decellularized scaffold, and forms a continuous cell layer on the surface of the scaffold through in vitro culture; S4: Once the cell layer is formed and cross-linked, the heart valve can be removed for further testing.

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