JP7251008B2 - Cell dispersion tool and cell dispersion device - Google Patents

Description

The present invention relates to cell dispersion tools and cell dispersion devices.

There are various techniques for culturing cells, but the obtained cells sometimes form aggregates in which a plurality of cells aggregate. In the case of subculturing this or when using it for regenerative medicine, pharmacological tests, etc., it is necessary to disperse the cell aggregates into individual cells. Proposed.

JP-A-4-232834 JP 2012-24061 A

Patent Documents 1 and 2 propose a mechanical device for dispersing cells, but the technique of Patent Document 1 has poor cell dispersion efficiency as pointed out in Patent Document 2, and Patent Document 2 The technique has a problem that the mechanism becomes complicated.

One of the objects of the present invention is to provide a cell dispersing device and a cell dispersing apparatus capable of efficiently dispersing a cell clump in which cells are aggregated into individual cells.

(1) The cell dispersion tool according to the present invention is
A tool for dispersing cell clumps into single cells, comprising:
a container containing a treatment liquid containing cell aggregates;
a rotator rotatably supported by the support shaft at the bottom of the recess formed with a recess into which the support shaft rising from the bottom surface of the container is inserted;
has
An air hole communicating between the inside and the outside of the recess is formed in the rotating body.

According to this, it is possible to provide a cell dispersing tool that facilitates attachment of the rotating body to the container.

(2) In the cell dispersion tool of this description,
The air hole may be arranged at a position higher than the surface of the treatment liquid when the rotating body is not rotating.

(3) In this cell dispersion tool,
The air hole may be formed in the bottom of the recess.

(4) In this cell dispersion tool,
The recess may be a tapered hole whose diameter increases toward the open end.

(5) A cell dispersion device according to the present invention has any of the cell dispersion tools described above.

FIG. 4 is a diagram for explaining a cell dispersion tool according to the embodiment; FIG. 4 is a diagram for explaining a cell dispersion procedure according to the present embodiment;

Embodiments to which the present invention is applied will be described below. However, the present invention is not limited to the following embodiments. That is, not all configurations described in the following embodiments are essential for the present invention. In addition, the present invention includes any combination of the following contents.

(1) Configuration of Cell Dispersion Device 1 First, the configuration of a cell dispersion device 1 according to an embodiment to which the present invention is applied will be described with reference to FIG.

A cell dispersion device 100 has a container 10 . The container 10 has a circular bottom surface and a cylindrical side surface, and serves to retain the processing liquid W inside. The container 10 has a support shaft 12 rising from the center of the bottom surface, and the tip of the support shaft 12 rotatably supports a rotating body 20 described later. The container 10 has a detachable cover 16. The cover 16 can prevent the inside of the container 10 from being contaminated and also prevent the processing liquid W from spilling out of the container 10. can.
The container 10 and the rotating body 20, which will be described later, are preferably made of a material that is inert to the treatment liquid W containing the cell aggregates and has resistance. Specifically, polycarbonate, polystyrene, acrylic resin, polyvinyl chloride, and the like can be applied.

The cell dispersing device 100 has a rotor 20 . The rotating body 20 is a member having a cylindrical outer shape and is accommodated in the container 10 . A recess 22 extending from the bottom surface to the top surface is formed in the rotating body 20 , and the rotating body 20 is housed in the container 10 with the support shaft 12 inserted into the recess 22 . Rotating body 20 is rotatably supported by spindle 12 (container 10 ) at bottom 24 of recess 22 . In this embodiment, a rotating shaft 26 formed in the bottom portion 24 of the recess 22 is configured to enter the recess at the tip of the support shaft 12 . In addition, in the present embodiment, the concave portion 22 is a tapered hole whose diameter increases toward the open end portion 28 .
The rotating body 20 is formed with an air hole 30 that communicates the inside and outside of the recess 22 . The air hole 30 can be configured as a through hole with one end opening to the inner surface of the recess 22 and the other end opening to the outer surface of the rotating body 20 . Although the air hole 30 is formed in the bottom portion 24 of the recess 22 in this embodiment, the present invention is not limited to this. For example, the air hole 30 can be arranged at a position higher than the surface of the processing liquid W when the rotating body 20 does not rotate.
In addition, in the present embodiment, the rotating body 20 has a convex portion formed on the upper surface thereof. As a result, since the rotating body 20 can be held, attachment and detachment to and from the container 10 is facilitated.

The cell dispersion device 100 has driving means 50 . The drive means 50 serves to rotate the rotating body 20 within the container 10 . In this embodiment, the drive means 50 has a drive magnet 52 and a motor 54 for rotating the drive magnet 52 . By rotating the driving magnet 52 , the driven magnet 56 provided on the rotating body 20 is driven to rotate the rotating body 20 .

According to the cell dispersing device 100 , by rotating the rotating body 20 within the container 10 , a centrifugal force acts on the processing solution W, and the processing solution W circulates inside the container 10 while being pressed against the inner surface of the container 10 . When the rotator 20 rotates within the container 10, a speed difference occurs between the inner wall surface of the container 10 and the outer surface of the rotator 20, so that a shearing force acts on the treatment liquid W circulating therebetween. . When this shearing force acts on the cell aggregates contained in the treatment liquid W, it becomes possible to disperse the cell aggregates into single cells.

(2) Cell Dispersion Procedure Next, a cell dispersion procedure using the cell dispersion apparatus 100 will be described with reference to FIGS. 2(a) to 2(c).
First, a cell dispersing tool 1 having a container 10 and a rotating body 20 is prepared. The cell dispersion tool 1 is preferably prepared in a sterilized state. Then, as shown in FIG. 2A, the processing liquid W is put into the container 10 . Then, as shown in FIG. 2B, the rotating body 20 is set in the container 10 and the lid 16 is attached. Thereafter, as shown in FIG. 2(c), the rotor 20 of the cell dispersion tool 1 is rotated to disperse the cell aggregates contained in the treatment liquid W into single cells. The drive conditions such as the rotation speed and rotation time of the rotating body 20 can be set to values suitable for the cells (cell aggregates) to be dispersed, and specifically can be determined experimentally. It is also possible to adopt a procedure in which the solution W is put into the container 10 with the rotating body 20 attached to the container 10 .

(3) Treatment liquid W
The treatment liquid W applicable to this embodiment is a solution containing cell aggregates.
Cell aggregates (cells) contained in the treatment liquid W are not particularly limited, and examples thereof include cells of animals, insects, plants, and the like. Animal cell sources include, but are not limited to, humans, monkeys, dogs, cats, rabbits, rats, nude mice, mice, guinea pigs, pigs, sheep, Chinese hamsters, cows, marmots, and African green monkeys. isn't it. In particular, the cell dispersing device 100 can be applied to cells such as pluripotent stem cells (iPS cells, ES cells, etc.), neural stem cells, and other stem cells, which have been considered to have poor dispersing efficiency in the prior art. Also, the method for obtaining cells to be processed by the cell dispersing device 100 is not particularly limited, and cells obtained by various methods can be used. For example, cell aggregates may be those produced by culturing, or tissue pieces (slices) derived from a living body.
Also, the solution constituting the treatment liquid W is not particularly limited. For example, a culture solution for culturing cells may be applied, and enzymes such as trypsin, protease and collagenase, and chelating agents such as EDTA may be added.

(4) Effect In the present embodiment, the rotor 20 of the cell dispersion device 100 (cell dispersion tool 1) is formed with the air holes 30 that communicate with the inside and outside of the recess 22. When attaching the 20 , air is discharged from the recess 22 and the processing liquid W enters the recess 22 . Therefore, the buoyant force that the rotating body 20 receives from the treatment liquid W can be reduced, and the rotating body 20 can be easily attached. Further, in this embodiment, the recessed portion 22 of the rotating body is a tapered hole whose diameter increases toward the open end portion 28 . According to this, the processing liquid W in the concave portion 22 is discharged from the opening end portion 28 by the centrifugal force acting on the processing liquid W as the rotating body 20 rotates, and the inner wall of the container 10 and the outer surface of the rotating body 20 are discharged. move between. At this time, since air enters the concave portion 22 through the air hole 30, it is possible to prevent part of the processing liquid W from remaining in the concave portion 22, and it is possible to apply a shearing force to the entire amount of the processing liquid W. In addition, it becomes difficult for air bubbles to enter the treatment liquid W in the step of dispersing the cell aggregates, so that the dispersion treatment can be performed without damaging the cells.

DESCRIPTION OF SYMBOLS 1... Cell dispersion tool 10... Container 12... Support shaft 16... Lid body 20... Rotating body 22... Recessed part 24... Bottom part 26... Rotating shaft 28... Opening end part 30... Air hole 50 ... drive means, 52 ... drive magnet, 54 ... motor, 56 ... driven magnet, 100 ... cell dispersing device, W ... treatment liquid

Claims (5)

A tool for dispersing cell clumps into single cells, comprising:
a container containing a treatment liquid containing cell aggregates;
a rotator rotatably supported by the support shaft at the bottom of the recess formed with a recess into which the support shaft rising from the bottom surface of the container is inserted;
has
The rotating body is configured to rotate around the support shaft by a predetermined driving means,
An air hole communicating between the inside and outside of the recess is formed in the rotating body ,
A cell dispersion tool in which the cell aggregates contained in the treatment liquid are dispersed into single cells by rotating the rotator around the spindle. The cell dispersion tool of claim 1, wherein
The cell dispersing tool, wherein the air hole is arranged at a position higher than the surface of the treatment liquid when the rotating body is not rotating. In the cell dispersion tool according to claim 1 or claim 2,
The cell dispersion tool, wherein the air hole is formed in the bottom of the recess. In the cell dispersion tool according to any one of claims 1 to 3,
The recess is a tapered hole with a diameter increasing toward the open end of the cell dispersing tool. A cell dispersion device comprising the cell dispersion tool according to any one of claims 1 to 4.

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