JP2010142137A - Cell-treating device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To collect a cell group from which unnecessary components are removed by efficiently cleaning the cells. <P>SOLUTION: The cell-treating device includes a container body 4 having a bottom part 4b gradually tapered to the tip, and rotatable around a prescribed axis by directing the bottom part 4b to the outside in the radial direction while storing a cell suspension A, a filter member 10 arranged at the position at which the interior of the container body 4 is divided in the depth direction and having a plurality of through holes having diameters allowing the cells contained in the cell suspension A to pass, a cleaning liquid-feeding tube 6 having an ejecting opening 6a arranged on the bottom part 4b side from the filter member 10, and a sucking tube 5 having a sucking opening 5a arranged at the position at the opposite side of the bottom part 4b so as to sandwitch the filter member 10. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a cell processing apparatus.

Conventionally, a centrifuge container containing a cell suspension obtained by decomposing adipose tissue and isolating adipose-derived cells is contained in the cell suspension by rotating it around an axis away from the centrifuge container. There is known a centrifugal separation device that separates components that are separated by specific gravity (see Patent Document 1).
The centrifuge container is formed in a substantially cylindrical shape with one end closed, and a component having a large specific gravity moves toward the closed end by rotating the closed end so that the closed end is directed radially outward. They are separated side by side in descending order of specific gravity.

International Publication No. 2005/012480 Pamphlet

  However, there is an inconvenience that the cell group separated at the bottom of the centrifuge container by centrifuging the cell suspension is formed into a solid pellet that is compressed by centrifugal force. That is, if the cell group is formed in a pellet shape, it becomes difficult to take out the cell group after centrifugation from the centrifuge container by suction. In addition, unnecessary components such as proteolytic enzymes and fats may be formed in the cell group during centrifugation, and may be formed into pellets, which makes it difficult to remove the unnecessary components.

  This invention is made | formed in view of the situation mentioned above, Comprising: It aims at providing the cell processing apparatus which can collect | recover the cell group from which the unnecessary component was removed by wash | cleaning a cell efficiently. .

In order to achieve the above object, the present invention provides the following means.
The present invention has a container body that has a bottom portion that gradually tapers toward the tip, accommodates a cell suspension, and is rotatable about a predetermined axis with the bottom portion directed radially outward, and the container body A filter member provided with a plurality of through holes having a diameter through which cells contained in the cell suspension can pass, and discharged toward the bottom side of the filter member. There is provided a cell treatment apparatus comprising a cleaning liquid supply pipe for disposing an outlet and a suction pipe for disposing a suction port at a position opposite to the bottom with the filter member interposed therebetween.

  According to the present invention, the cell suspension is contained in the container body, and the container body is contained in the cell suspension by rotating the container body about a predetermined axis with its bottom portion radially outward. The components are separated by specific gravity, a group of cells having a large specific gravity is collected near the bottom of the container body, and a supernatant such as plasma is collected in the upper layer. The cell group is pressed into a pellet by centrifugal force.

In this state, when suction is performed from the suction port of the suction tube, the supernatant is sucked from the suction port, and a pellet-like cell group can be left in the vicinity of the bottom. Then, when the cleaning liquid is discharged from the discharge port of the cleaning liquid discharge pipe, the pellet-like cell group is collapsed by the momentum of the cleaning liquid, and unnecessary components taken into the inside can be released.
In this case, the collapsed cell group is finely separated when it is allowed to pass through the through holes by the momentum of the cleaning liquid passing through the filter member. As a result, it is possible to prevent inconvenience that the cell group remains as a large lump.

In the said invention, the said filter member may be formed in flat form with the elastic material which can be elastically deformed with a centrifugal force, and the said through-hole may be penetrated and formed in the plate | board thickness direction.
By doing in this way, since the filter member is elastically deformed by rotating the container body and applying a centrifugal force to the filter member, the aperture of the through hole provided in the filter member can be changed. . That is, when the cleaning liquid is discharged from the discharge port in a state where the diameter of the through hole of the filter member is enlarged, the filter member is finely separated when being passed through the through hole by the force of the cleaning liquid passing through the filter member. On the other hand, the supernatant can be removed from the cell group without rolling up the cells by elastically deforming the filter member so as to reduce the diameter of the through hole of the filter member and sucking the supernatant from the suction port. . As a result, the cell yield can be improved.

  Moreover, in the said invention, when the said filter member is receiving the centrifugal force more than predetermined value, when the said filter member is expanded to the aperture which the said cell can pass and centrifugal force becomes smaller than predetermined value, The diameter may be reduced to be smaller than the cell.

  According to the present invention, the cell group from which unnecessary components are removed by efficiently washing the cells can be collected.

A cell treatment apparatus 1 according to an embodiment of the present invention will be described below with reference to the drawings.
As shown in FIG. 1, the cell treatment apparatus 1 according to the present embodiment includes a container body 4 that is horizontally rotated around a vertical axis 3 by a motor 2, and is fixed to the container body 4. A supernatant suction pipe 5 for sucking the separated supernatant and discharging it out of the container body 4, a cleaning liquid supply pipe 6 fixed to the container body 4 and supplying a cleaning liquid into the container body 4, a fixed pipe 7, A rotary joint 8 that connects the supernatant suction pipe 5 and the washing liquid supply pipe 6 and a peristaltic pump 9 that feeds and discharges the liquid in the fixed pipe are provided.

As shown in FIG. 2A, the container body 4 has a cylindrical side wall 4a and a bottom surface (bottom) 4b disposed at a position where one end of the side wall 4a is closed. The bottom surface 4b is formed in a tapered inner surface shape that tapers gradually toward the tip.
In the container main body 4, a flat plate-like filter member 10 is provided so as to divide the space in the container main body 4 into two in the depth direction.

  The filter member 10 is made of an elastic material such as rubber, for example. The filter member 10 is provided with a large number of through holes 11 penetrating in the plate thickness direction. In the state where the filter member 10 is formed in a flat plate shape, the through hole 11 has a sufficiently small or almost closed diameter as shown in FIG.

On the other hand, as shown in FIG. 3A, in the state in which the filter member 10 is elastically deformed in the plate thickness direction so that the central portion is raised with respect to the peripheral portion, the elasticity constituting the filter member 10 As a result of the elongation of the material, the aperture 11 also has an enlarged diameter as shown in FIG. The outer peripheral edge of the filter member 10 is fixed to the inner surface of the container body 4. When a centrifugal force acts on the container body 4, the vicinity of the center portion of the filter member 10 moves in the direction toward the bottom surface 4 b of the container body 4 by the centrifugal force so that the center portion rises with respect to the peripheral edge portion. It can be elastically deformed.
In the present embodiment, the through hole 11 of the filter member 10 is expanded to a diameter that allows cells to pass through in an expanded state, for example, a diameter of 100 μm or more.

The supernatant suction tube 5 has a suction port 5a disposed on the side opposite to the bottom surface 4b of the container body 4 with the filter member 10 interposed therebetween.
On the other hand, the discharge port 6 a of the cleaning liquid supply pipe 6 is disposed near the bottom surface between the filter member 10 and the bottom surface 4 b of the container body 4.

The operation of the cell processing apparatus 1 according to the present embodiment configured as described above will be described below.
In order to take out the cell group C from the cell suspension A using the cell processing apparatus 1 according to the present embodiment, the cell suspension A is accommodated in the container body 4 and the motor 2 is operated to operate the container body. Rotate 4 horizontally. When the rotational speed increases and the centrifugal force increases, the container body 4 is swung so that the tapered bottom surface 4b is directed outward in the radial direction, and is rotated horizontally in that state.

  As shown in FIG. 2, the cell suspension A is centrifuged into a supernatant B and a cell group C according to the specific gravity. The cell group C moves toward the portion of the container body 4 where the centrifugal force is the largest, that is, toward the bottom surface 4b arranged at the outermost side in the radial direction. At this time, the elastic material constituting the filter member 10 is elastically deformed by centrifugal force, and the through-hole 11 is enlarged, so that the cells contained in the cell suspension A are separated by the filter member 10. Without being blocked, it passes through the through-hole 11 and is collected in the vicinity of the bottom surface 4b, rolls on the bottom surface 4b having a tapered inner surface, and is deposited in a pellet form at the tip. At this time, it is formed into a pellet in a state where unnecessary components such as digestive enzyme solution and dust contained in the cell suspension A are taken up between cells.

  In this state, when the rotational speed of the motor 2 is reduced, the elastic deformation of the filter member 10 that has been elastically deformed by the centrifugal force is restored and returns to a substantially flat plate shape. Therefore, the through hole 11 provided in the filter member 10 Is reduced, and the through hole 11 is closed. As a result, the inside of the container body 4 is divided into two in the longitudinal direction. When the perstaltic pump 9 is operated in this state and the supernatant B is sucked from the suction port 5a of the supernatant suction tube 5, the supernatant B is discharged to the outside of the container body 4 through the suction port 5a, the rotary joint 8 and the fixed pipe 7.

  In this case, according to the present embodiment, when the supernatant B is sucked, the centrifugal force is reduced to close the through hole 11 of the filter member 10, so that the flow of the supernatant B due to the suction of the supernatant B is It becomes difficult to be transmitted to the pellet-like cell group C. Accordingly, there is an advantage that the cell yield can be improved by preventing the disadvantage that the cells flow up from the cell group C due to the flow of the supernatant B and are discharged from the suction port 5a.

  Further, when supplying the cleaning liquid D from the cleaning liquid supply pipe 6, the cleaning liquid D is discharged from the discharge port 6a in a state where the rotational speed of the motor 2 is increased and the centrifugal force is increased. Thereby, the filter member 10 is elastically deformed by the centrifugal force, the through hole 11 is enlarged, and the cell suspension A ′ can pass through the filter member.

  That is, when the pellet-like cell group C is unwound and resuspended by the flow of the cleaning liquid D discharged from the discharge port 6a, the cell group C passes through the enlarged through hole 11 of the filter member 10. To be forced into. Since the through-hole 11 of the filter member 10 is enlarged to such a size that a cell can pass through, a cell mass in which a plurality of cells are gathered cannot pass through but is separated by the through-hole 11. Then, the cells in a separated state are allowed to pass through the through holes. As a result, it is possible to obtain a cell suspension A ′ resuspended in a state where the cells are more finely separated.

As a result, the pellet-shaped cell group C that is wound with the unnecessary component and consolidated is resuspended by the washing liquid D, so that the unnecessary component is released from the cell group C.
Then, by rotating the container body 4 horizontally again and centrifuging the resuspended cell suspension A ′, the cell group C from which unnecessary components have been removed is formed at the tip of the bottom surface 4b in a pellet form. be able to.

  Then, the cell group C consolidated into a pellet can be accommodated by discharging the supernatant B in the container body 4 by operating the supernatant suction tube 5 with the centrifugal force reduced again. Thereafter, the cell group C accumulated at the tip of the container body 4 can be taken out by operating the cell suction tube that also serves as the cleaning liquid supply tube 6.

Thus, according to the cell processing apparatus 1 which concerns on this embodiment, by letting the filter member 10 pass, the pellet-shaped cell group C is decomposed | disassembled more finely, an impurity is discharge | released, and the cell group C of a healthy state is obtained. There is an advantage that it can be recovered.
In addition, by aspirating the supernatant B in a state where the centrifugal force is reduced, the influence of the flow of the supernatant B is prevented from being exerted on the cell group C. Therefore, it is possible to prevent the occurrence of inconvenience that is sucked. Thereby, the collection rate of the cell group C finally obtained can be improved.

  In this embodiment, examples of the elastic material such as rubber that is the material of the filter member 10 include silicone resin, butadiene rubber, styrene-butadiene rubber, isoprene rubber, ethylene-propylene rubber, butyl rubber, and acrylonitrile-butadiene rubber. Carboxonitrile rubber, acrylic rubber, hydrogenated nitrile rubber, urethane rubber, fluorine rubber, elastomer such as thermoplastic rubber, and natural rubber. These may be used alone or in combination of two or more. Among these, a silicone resin or the like is preferable in terms of good biocompatibility.

Further, the filter member 10 may be formed of a flat plate member that is not elastically deformed so much. In this case, the filter member 10 only needs to have a diameter that allows cells to pass through.
In this way, it is impossible to prevent the cell group C from being affected by the suction of the supernatant B. However, since the cell group C is dissociated finely when passing through the through holes 11 of the filter member 10, the cells A foreign substance sandwiched between them can be sufficiently released to obtain a healthy cell group C.

  Further, in this embodiment, the cell suspension A accommodated in the container body 4 is not particularly limited as long as it is a suspension containing living cells, but for example, derived from fat containing cells derived from fat. A cell suspension or the like is preferred. Here, examples of the adipose-derived cells include vascular endothelial cells, fibroblasts, hematopoietic stem cells, hematopoietic cells, adipose-derived stem cells, and the like. This stem cell is a cell collected from adipose tissue for the purpose of cell therapy or the like, and refers to a cell having multipotency and self-renewal ability.

It is a front view which shows the cell processing apparatus which concerns on one Embodiment of this invention. In the cell processing apparatus of FIG. 1, (a) The longitudinal cross-sectional view which shows the structure inside a container main body in the state which reduced the centrifugal force, (b) The top view which shows the state of a filter. In the cell processing apparatus of FIG. 1, (a) The longitudinal cross-sectional view which shows the structure inside a container main body in the state which increased the centrifugal force, (b) The top view which shows the state of a filter.

Explanation of symbols

A Cell suspension C Cell group 1 Cell processing device 4 Container body 4b Bottom (bottom)
DESCRIPTION OF SYMBOLS 5 Suction pipe 5a Suction port 6 Cleaning liquid supply pipe 6a Discharge port 10 Filter member 11 Through-hole

Claims (3)

A container body that has a bottom that gradually tapers toward the tip, accommodates a cell suspension, and is rotatable about a predetermined axis with the bottom facing radially outward;
A filter member provided with a plurality of through-holes having a diameter that allows cells contained in the cell suspension to pass through, which are arranged at positions that divide the container body in the depth direction;
A cleaning liquid supply pipe that disposes a discharge port on the bottom side of the filter member;
A cell treatment apparatus comprising: a suction tube having a suction port disposed at a position opposite to the bottom with the filter member interposed therebetween. The filter member is formed in a flat plate shape by an elastic material that can be elastically deformed by centrifugal force,
The cell treatment device according to claim 1, wherein the through-hole is formed so as to penetrate the filter member in a plate thickness direction.   The through hole is expanded to a diameter through which the cells can pass when the filter member receives a centrifugal force greater than or equal to a predetermined value. When the centrifugal force becomes smaller than a predetermined value, the through hole has a smaller diameter than the cells. The cell processing apparatus according to claim 2, which is reduced.

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