Multi-cavity cell bag
By designing a multi-chamber cell bag and utilizing the structure of separate chambers and a total chamber, the problem of cell spheroid density reduction after high-density culture was solved, the cell spheroid formation rate and culture quality were improved, and cell spheroid size uniformity was achieved.
Patent Information
- Application Number
- CN202511239764.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-01
- Publication Date
- 2025-11-18
AI Technical Summary
In existing technologies, cell spheres tend to fuse together during high-density culture, leading to size inhomogeneity. There is a lack of cell bags that can reduce density after initial high-density culture.
Design a multi-chamber cell bag comprising sub-chambers and a main chamber connected by a controllable channel. The sub-chambers are used for high-density culture, while the main chamber is used for low-density culture. It is equipped with multiple sample inlet, liquid outlet, and aeration ports to meet the environmental requirements of different growth stages of the cell spheres.
By designing separate chambers and a total chamber, the cell spheroid formation rate and culture quality were improved, the size uniformity of the cell spheroids was maintained, and high-quality experimental materials were provided for cell spheroid-related research.
Smart Images
Figure CN120966634A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to a cell bag, in particular to a multi-cavity cell bag. BACKGROUND
[0002] In the culture technology of cell spheres, cells and culture medium are usually injected into the same cell bag for culture, and the cell spheres are taken out from the cell bag after being cultured into spheres.
[0003] The cell spheres need to be cultured at a high density in the initial stage, and the cells form a cell sphere structure through interaction, and each mesenchymal stem cell makes the cell sphere structure gradually stable through the secretion of extracellular matrix proteins. However, due to the high-density culture at the beginning, the number of sphere units in the unit volume is high, which leads to the fact that the spheres are very easy to fuse with each other after being formed, and seriously affects the uniformity of the size of the cell spheres. The culture system after the formation of the sphere units can reduce the content of the spheres in the unit volume, reduce the direct fusion of the cell spheres, and thus the uniformity of the size can be maintained. However, there is no such cell bag in the prior art that can realize the above-mentioned related functions. SUMMARY
[0004] The purpose of the present application is to provide a multi-cavity cell bag to solve the problems raised in the background art.
[0005] To achieve the above-mentioned purpose, the present application provides the following technical solutions:
[0006] A multi-cavity cell bag is placed flat on a platform and used for culturing cell spheres, comprising a cell bag body, a plurality of sub-cavity bodies for culturing cell spheres and a total cavity body for collecting the culture and reducing the density of the cell spheres are arranged in the cell bag body, the sub-cavity bodies and the total cavity body are connected through channels with controllable opening and closing states, a first sampling port for adding cells and culture medium into the sub-cavity body and a first liquid discharge port for discharging the waste liquid of the sub-cavity body culture are arranged on each sub-cavity body, a second sampling port for adding culture medium into the total cavity body, a sampling port for taking out cell spheres and a second liquid discharge port for discharging the waste liquid of the total cavity body culture are arranged on the total cavity body, and a first gas exchange port and a second gas exchange port for exchanging gas during the cell sphere culture are arranged on the sub-cavity body and the total cavity body, respectively.
[0007] Compared with the prior art, the present application has the following beneficial effects:
[0008] The present application first induces high-density cell spheres in the sub-cavity body, and then collects them in the total cavity body for low-density culture, which meets the needs of the cell spheres in different growth stages and is beneficial to improve the formation rate and culture quality of the cell spheres, and provides high-quality experimental materials for the related research of the cell spheres. BRIEF DESCRIPTION OF DRAWINGS
[0009] Figure 1 It is a top view of a multi-cavity cell bag.
[0010] Figure 2 is a bottom view of a multi-chamber cell bag. DETAILED DESCRIPTION
[0011] The technical solutions in the embodiments of the present application will be clearly and completely described in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the present application.
[0012] In the embodiments of the present application, a multi-chamber cell bag is placed on a platform and used for culturing cell spheres, especially for culturing mesenchymal stem cell spheres, which comprises a cell bag body 1, a plurality of sub-chamber bodies 2 for culturing cell spheres and a total-chamber body 3 for collecting culture and reducing the density of cell spheres are arranged in the cell bag body 1, the sub-chamber bodies 2 and the total-chamber body 3 are connected through channels 4 with controllable on-off state, a first sample inlet 5 for adding cells and culture medium into the sub-chamber bodies 2 and a first liquid discharge port 9 for discharging culture waste liquid of the sub-chamber bodies 2 are arranged on each sub-chamber body 2, a second sample inlet 7 for adding culture medium into the total-chamber body 3, a sampling port 10 for taking out cell spheres and a second liquid discharge port 11 for discharging culture waste liquid of the total-chamber body 3 are arranged on the total-chamber body 3, and a first gas exchange port 6 and a second gas exchange port 8 for exchanging gas during the culture of cell spheres are arranged on the sub-chamber bodies 2 and the total-chamber body 3, respectively.
[0013] As shown in Figs. 1 and 2, the lower part of the cell bag body 1 is a large total-chamber body 3, and the upper part of the cell bag body 1 is a plurality of parallel sub-chamber bodies 2. In addition to this form, the sub-chamber bodies 2 can be distributed around the total-chamber body 3, and can also be distributed in a three-dimensional form outside the total-chamber body 3. Other forms are not listed here. Figure 1 Figure 2 As shown in Figs. 1 and 2, the lower part of the cell bag body 1 is a large total-chamber body 3, and the upper part of the cell bag body 1 is a plurality of parallel sub-chamber bodies 2. In addition to this form, the sub-chamber bodies 2 can be distributed around the total-chamber body 3, and can also be distributed in a three-dimensional form outside the total-chamber body 3. Other forms are not listed here.
[0014] The capacity of the total-chamber body 3 is greater than the sum of the capacities of all the sub-chamber bodies 2, and is preferably 1.5-3 times the sum of the capacities of the sub-chamber bodies 2. The density of cell spheres in the sub-chamber bodies 2 is relatively high, and the density of cell spheres can be reduced after the cell spheres that have been preliminarily cultured enter the total-chamber body 3.
[0015] A filter screen (not marked in the figure) is fixed in each of the first liquid discharge port 9 and the second liquid discharge port 11 to avoid the discharge of cell spheres when waste liquid is discharged, so that the cell spheres are retained in the sub-chamber body 2 / total-chamber body 3 when the waste liquid is discharged.
[0016] The mesh diameter of the filter screen is not greater than 0.5 μm. Since the single cell size of mesenchymal stem cells is usually greater than 10 μm, which is greater than the mesh diameter, when the waste liquid is drained (e.g. when the waste liquid in the sub-cavity 2 or the total cavity 3 is replaced), the waste liquid can flow out through the filter screen, and the mesenchymal stem cells and cell spheres are retained, thereby effectively avoiding the loss of cells with the waste liquid.
[0017] A valve (not shown in the figure) capable of controlling the on-off state of the passage is arranged in the passage 4 between the total cavity 3 and the sub-cavity 2. The passage 4 is a hard pipeline. When the cells are cultured into spheres in the sub-cavity 2, the valve is in a closed state. After the preliminary culture is completed, the culture bag is erected (the sub-cavity 2 is above the total cavity 3), the cell spheres and the culture liquid are gathered around the valve, the valve is opened, and the cell spheres and the culture liquid in the sub-cavity 2 filled with gas are pressed into the total cavity 3 by the gas pressure in the sub-cavity 2. After the cell spheres and the culture liquid in the sub-cavity 2 are all introduced into the total cavity 3, the valve is closed.
[0018] In the embodiment, the connection of the first sampling port 5, the first gas exchange port 6, the second sampling port 7 and the second gas exchange port 8 with the cell bag body 1 is located on the upper surface of the cell bag body 1; and the connection of the first drainage port 9, the sampling port 10 and the second drainage port 11 with the cell bag body 1 is located on the lower surface of the cell bag body 1.
[0019] A sealing member is arranged on each of the first sampling port 5, the first gas exchange port 6, the second sampling port 7, the second gas exchange port 8, the first drainage port 9, the sampling port 10 and the second drainage port 11. Preferably, the sealing members on the first sampling port 5, the second sampling port 7, the first drainage port 9, the sampling port 10 and the second drainage port 11 are sealing nuts; and the sealing members on the first gas exchange port 6 and the second gas exchange port 8 are stop-type reversing valves.
[0020] It should be noted that the cell bag body in the present application is made of a common transparent material such as medical polyester. When the cell spheres are cultured, the cells and the culture medium are first injected into each sub-cavity 2 for preliminary high-cell-density culture. After the cell spheres that have undergone the preliminary culture enter the total cavity 3, the density of the cell spheres can be reduced. During the culture process, the culture waste liquid needs to be drained from the first drainage port 9 / second drainage port 11 and the new culture medium needs to be injected from the first sampling port 5 / second sampling port 7, regardless of the sub-cavity 2 or the total cavity 3. After the culture is completed, the cell spheres in the total cavity 3 are taken out from the sampling port 10.
[0021] Although the present application has been described in detail with reference to the foregoing embodiments, the technical solutions recorded in the foregoing embodiments can be modified or some of the technical features can be replaced by equivalent ones by those skilled in the art, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application, and the contents not described in detail in the specification are all the prior art known by those skilled in the art.
Claims
1. A multi-chamber cell bag, placed flat on a platform, for culturing cell spheres, characterized in that, The system includes a cell bag body (1), which contains multiple sub-cavities (2) for culturing cell spheres and a total cavity (3) for collecting culture and reducing cell sphere density. The sub-cavities (2) and the total cavity (3) are connected by a channel (4) with controllable on / off state. Each sub-cavity (2) has a first inlet (5) for adding cells and culture medium into the sub-cavity (2) and a first drain outlet (9) for discharging culture waste liquid from the sub-cavity (2). The total cavity (3) has a second inlet (7) for adding culture medium into the total cavity (3), a sampling port (10) for removing cell spheres, and a second drain outlet (11) for discharging culture waste liquid from the total cavity (3). The sub-cavities (2) and the total cavity (3) have a first gas exchange port (6) and a second gas exchange port (8) for exchanging gases during cell sphere culture, respectively.
2. The multi-chamber cell bag according to claim 1, characterized in that, The total capacity of the main cavity (3) is greater than the sum of the capacities of all the sub-cavities (2).
3. The multi-chamber cell bag according to claim 1, characterized in that, Both the first drain port (9) and the second drain port (11) are fitted with filters to prevent cell spheres from being discharged during waste liquid discharge.
4. A multi-chamber cell bag according to claim 3, characterized in that, The mesh diameter of the filter screen is no greater than 0.5 μm.
5. A multi-chamber cell bag according to claim 1, characterized in that, The passage (4) between the main cavity (3) and the sub-cavities (2) is equipped with a valve that can control the opening and closing of the passage.
6. A multi-chamber cell bag according to claim 1, characterized in that, The connection points of the first sample inlet (5), the first air exchange port (6), the second sample inlet (7), and the second air exchange port (8) with the cell bag body (1) are located on the upper surface of the cell bag body (1); the connection points of the first drain port (9), the sampling port (10), and the second drain port (11) with the cell bag body (1) are located on the lower surface of the cell bag body (1).