Microporous stem cell balling device

Abstract

The invention discloses a microporous stem cell balling device, and relates to the technical field of cell ball culture, the microporous stem cell balling device comprises a culture zone for culture, the culture zone is provided with a plurality of culture holes for culturing mesenchymal stem cell balls; when the cell balls are cultured, a culture solution with mesenchymal stem cells is arranged in the culture holes for culture and growth, and the cells are taken out until the cells are cultured into balls in the micropores. According to the scheme design, when the mesenchymal stem cell spheres are cultured, a culture solution does not need to be stirred or vibrated or shaken, and culture is more convenient; and the sizes of the formed cell balls are based on the sizes of the culture holes, so that the formed cell balls are more uniform and consistent in size.

Description

Technical Field

[0001] The invention relates to the technical field of cell sphere culture, in particular to a microporous stem cell sphere forming device. Background Art

[0002] In the field of bioengineering, the cultivation of mesenchymal stem cell spheres is of great significance for regenerative medicine, drug development, and many other fields. However, the current devices used to culture mesenchymal stem cell spheres have many problems that need to be solved.

[0003] Traditional culture devices often require stirring, oscillating, or shaking the culture medium when culturing mesenchymal stem cell spheres. This not only increases the complexity of the culture process, but may also damage the cells due to improper operation, affecting the normal formation of cell spheres, and the size consistency of the formed cell spheres is poor. Summary of the Invention

[0004] The purpose of the present invention is to provide a microporous stem cell spheroidization device to solve the problems raised in the above background technology.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] A microporous stem cell sphere formation device is placed in an incubator for culturing mesenchymal stem cell spheres. It includes a culture carrier with a carrying surface. The surface of the culture carrier is provided with multiple culture wells arranged in an array. When culture fluid containing mesenchymal stem cells is injected into the culture wells, the cells spontaneously aggregate into spheres in the culture wells.

[0007] Preferably, the culture carrier is a flexible belt, and the culture holes are distributed on the surface of the belt.

[0008] Preferably, the culture well is a hemispherical well with a diameter of 0.1-0.5 mm.

[0009] Preferably, the culture belt is a linear belt structure or a closed-loop belt structure connected end to end.

[0010] Preferably, the culture belt is a closed-loop belt structure connected end to end, and the culture holes are located on the outer surface of the culture belt.

[0011] Preferably, brackets for installing the culture belt are provided on both sides of the culture belt, and multiple parallel transmission mechanisms are provided between the brackets. The culture belt passes around all the transmission mechanisms in turn to form a closed-loop motion path. When any one of the transmission mechanisms moves, the culture belt and all the transmission mechanisms are driven to move synchronously.

[0012] Preferably, the transmission mechanism includes a transmission shaft and connecting sleeves arranged at both ends of the transmission shaft. The two ends of the transmission shaft are rotatably connected to the brackets arranged on both sides of the culture belt through the connecting sleeves. The two side parts of the culture belt are overlapped on the connecting sleeves. The diameter of the transmission shaft is smaller than the outer diameter of the connecting sleeve, so that the culture belt has no contact with the transmission shaft during movement.

[0013] Preferably, a rib is fixed on the connecting sleeve, and the spacing between the ribs at both ends of the transmission shaft is not less than the width of the culture belt, which is used to limit the culture belt from shaking too much during operation.

[0014] Preferably, the method further comprises a taking-out mechanism for taking out the cultured mesenchymal stem cell spheres on the culture belt.

[0015] Preferably, the culture belt is provided with an air hole penetrating the culture belt at a position corresponding to each culture hole.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention adopts the above-mentioned design scheme, and when culturing mesenchymal stem cell spheres, it is no longer necessary to stir, oscillate or shake the culture medium, making the culture more convenient. Moreover, the size of the formed cell spheres is based on the size of the culture wells, so the formed cell spheres are more uniform and of consistent size. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a schematic diagram of the structure of a microporous stem cell spheroidization device.

[0019] Figure 2 This is a schematic diagram of the structure of the transmission mechanism in a microporous stem cell spheroidization device.

[0020] Figure 3 Schematic diagram of the structure of the removal device in a microporous stem cell spheroidization device Figure 1 .

[0021] Figure 4 Schematic diagram of the structure of the removal device in a microporous stem cell spheroidization device Figure 2 .

[0022] Figure 5 Schematic diagram of the structure of the removal device in a microporous stem cell spheroidization device Figure 3 . DETAILED DESCRIPTION

[0023] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0024] See also Figures 1 to 5 In an embodiment of the present invention, a microporous stem cell sphere forming device is placed in an incubator for culturing mesenchymal stem cell spheres, including a culture carrier 1 having a carrying surface, and the surface of the culture carrier 1 is provided with a plurality of culture wells 3 arranged in an array. When a culture medium containing mesenchymal stem cells is injected into the culture wells 3, the cells spontaneously aggregate into spheres in the culture wells 3.

[0025] The present invention further provides a liquid distribution pipe 6 above the culture belt 1, and a plurality of liquid distribution nozzles 7 are provided on the liquid distribution pipe 6. When distributing liquid, the liquid distribution pipe 6 is connected to the cell culture liquid supply source, and the culture liquid is arranged into the culture hole 3 through the liquid distribution nozzle 7. It should be noted that the liquid distribution nozzle 7 can use a nozzle with a solenoid valve, and the existing electronic control technology is used to control the liquid spraying operation of the liquid distribution nozzle 7. As an existing technology, this technology will not be described in detail here.

[0026] Through the above-mentioned design, the present invention eliminates the need to stir, oscillate, or shake the culture medium when culturing mesenchymal stem cell spheres, making the culture more convenient. Furthermore, the size of the formed cell spheres is based on the size of the culture wells 3, so the formed cell spheres are more uniform and of uniform size.

[0027] The culture carrier 1 is a flexible belt, and the culture holes 3 are distributed on the surface of the belt.

[0028] The culture well 3 is a hemispherical well with a diameter of 0.1-0.5 mm.

[0029] The culture belt 1 is a linear belt structure or a closed-loop belt structure connected end to end. In the present application, the belt structure connected end to end is preferred, and the culture holes 3 are located on the outer surface of the culture belt 1 .

[0030] Brackets 4 for installing the culture belt 1 are provided on both sides of the culture belt 1, and multiple parallel transmission mechanisms 5 are provided between the brackets 4. The culture belt 1 passes around all the transmission mechanisms 5 in sequence to form a closed-loop motion path. When any one of the transmission mechanisms 5 moves, the culture belt 1 and all the transmission mechanisms 5 are driven to move synchronously; when driving the culture belt 1 to move, a drive motor (not shown in the figure) can be installed on the bracket 4 to drive any one of the transmission mechanisms 5 to rotate, thereby driving the culture belt 1 to move.

[0031] After the liquid is applied, the culture wells 3 with the liquid applied will face downward as the culture belt 1 moves. At this time, the culture liquid can stay in the culture wells 3 due to the surface tension of the culture liquid.

[0032] The transmission mechanism 5 includes a transmission shaft 53 and connecting sleeves 51 arranged at both ends of the transmission shaft 53. The two ends of the transmission shaft 53 are rotatably connected to the brackets 4 arranged on both sides of the culture belt 1 through the connecting sleeves 51. The two side portions of the culture belt 1 are overlapped on the connecting sleeves 51. The diameter of the transmission shaft 53 is smaller than the outer diameter of the connecting sleeve 51, so that the culture belt 1 has no contact with the transmission shaft 53 during movement, and does not affect the culture of cell spheres.

[0033] A rib 52 is fixed on the connecting sleeve 51 , and the spacing between the ribs at both ends of the transmission shaft 53 is not less than the width of the culture belt 1 , which is used to limit the culture belt 1 from excessive shaking during operation and ensure the operation stability of the culture belt 1 .

[0034] A matching driving structure is provided between the connecting sleeve 51 and the culture belt 1. When either the connecting sleeve 51 or the culture belt 1 is in operation, the other component is driven to operate.

[0035] The driving structure is a gear arranged on the circumference of the outer surface of the connecting sleeve 51 and a gear hole 2 opened on both sides of the culture belt 1. The cooperation between the gear and the gear hole 2 enables the culture belt 1 and the connecting sleeve 51 to move synchronously.

[0036] The device also includes a taking-out mechanism for taking out the mesenchymal stem cell spheres cultured on the culture belt 1 .

[0037] An air hole 8 penetrating the culture belt 1 is provided at a position corresponding to each culture hole 3 , and the inner diameter of the air hole 8 is smaller than the inner diameter of the culture hole 3 .

[0038] The removal mechanism is a pneumatic sampling mechanism, which includes the following two implementation methods:

[0039] 1. The removal mechanism includes a sampling bracket 9 arranged above or below any section of the culture belt 1. A plurality of sampling cylinders 10 corresponding to the culture wells 3 are fixed on the sampling bracket 9. The sampling ports of the sampling cylinders 10 face the culture wells 3, and the sampling cylinders 10 are connected to a negative pressure source. The cell spheres formed in the culture wells 3 are sucked out by the negative pressure.

[0040] 2. The removal mechanism includes an air blowing tube 12 arranged above any section of the culture belt 1 and a collection pool 11 arranged below the culture belt 1 at the corresponding position. The culture holes 3 of the culture belt 1 in this section are located on the lower surface of the culture belt 1. The air blowing tube 12 is connected to an external air source and is provided with multiple downward-facing air outlets. When sampling, the gas blows the formed cell spheres out through the air holes 8 and drops them into the collection pool 11.

[0041] To culture spheroids, the device is placed in a standard cell culture incubator (e.g., 37°C, 5% CO2). First, a mesenchymal stem cell suspension is precisely injected into the culture wells 3 via the liquid distribution tube 6. Subsequently, the culture belt 1, driven by a transmission mechanism, slowly rotates (at a speed of 0.1-0.5 mm / min), allowing the cells to self-aggregate in the culture wells 3 in a static environment. After a culture period of 3-7 days, the cells form spheroids within the microwells, which are then removed using a pneumatic sampling mechanism.

[0042] Although the embodiments have been described in detail, modifications by those skilled in the art, such as adjusting the hole size or drive method, without departing from the spirit of the present invention (stir-free and high uniformity), are within the scope of the present invention. Portions not described in detail (such as the electronic control system) are implemented using known techniques.

Claims

1. A microporous stem cell sphere forming device placed in an incubator for culturing mesenchymal stem cell spheres, characterized in that: The invention comprises a culture carrier (1) having a carrying surface, wherein the surface of the culture carrier (1) is provided with a plurality of culture wells (3) arranged in an array. When a culture fluid containing mesenchymal stem cells is injected into the culture wells (3), the cells spontaneously aggregate into spheres in the culture wells (3).

2. The microporous stem cell spheroidization device according to claim 1, characterized in that: The culture carrier (1) is a flexible belt, and the culture holes (3) are distributed on the surface of the belt.

3. A microporous stem cell spheroidization device according to claim 1 or 2, characterized in that: The culture hole (3) is a hemispherical hole with a diameter of 0.1-0.5 mm.

4. A microporous stem cell spheroidization device according to claim 1 or 2, characterized in that: The culture belt (1) is a linear belt structure or a closed-loop belt structure connected end to end.

5. The microporous stem cell spheroidization device according to claim 4, characterized in that: The culture belt (1) is a belt-type structure with a closed loop connected end to end, and the culture holes (3) are located on the outer surface of the culture belt (1).

6. The microporous stem cell spheroidization device according to claim 1 or 5, characterized in that: Brackets (4) for mounting the culture belt (1) are provided on both sides of the culture belt (1), and a plurality of transmission mechanisms (5) arranged in parallel are provided between the brackets (4). The culture belt (1) passes through all the transmission mechanisms (5) in sequence to form a closed-loop motion path. When any one of the transmission mechanisms (5) moves, the culture belt (1) and all the transmission mechanisms (5) are driven to move synchronously.

7. The microporous stem cell spheroidization device according to claim 6, characterized in that: The transmission mechanism (5) comprises a transmission shaft (53) and connecting sleeves (51) arranged at both ends of the transmission shaft (53). The two ends of the transmission shaft (53) are rotatably connected to the brackets (4) arranged at both sides of the culture belt (1) through the connecting sleeves (51). The two side portions of the culture belt (1) overlap on the connecting sleeves (51). The diameter of the transmission shaft (53) is smaller than the outer diameter of the connecting sleeves (51), so that the culture belt (1) has no contact with the transmission shaft (53) during movement.

8. The microporous stem cell spheroidization device according to claim 7, characterized in that: A rib (52) is fixed on the connecting sleeve (51), and the spacing between the ribs at both ends of the transmission shaft (53) is not less than the width of the culture belt (1), so as to limit the culture belt (1) from excessive shaking during operation.

9. The microporous stem cell spheroidization device according to claim 1, characterized in that: It also includes a taking-out mechanism for taking out the cultured mesenchymal stem cell balls on the culture belt (1).

10. The microporous stem cell spheroidization device according to claim 1 or 9, characterized in that: The culture belt (1) is provided with an air hole at a position corresponding to each culture hole (3) and passing through the culture belt (1).

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