JP2010124784A - Cell-cleansing system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To swiftly feed cells with a bubble-free cleansing liquid while preventing a bubble-containing cleansing liquid from being fed. <P>SOLUTION: A cell-cleansing system 1 is provided, including: a cell container 2; a feed pipe 7 to feed a cleansing liquid into the container 2; a pump 8 to feed the cleansing liquid in the feed pipe 7 toward the container 2; a reservoir 10, that is set midway of the feed pipe 7, so as to extend above the feed pipe 7 and composed of a recessed part for catching bubbles in the cleansing liquid; a sensor 9 installed in the feed pipe 7 between the pump 8 and the reservoir 10 and functioning to detect bubbles flowing in the feed pipe 7; and a controller 11 to make a control so as to reduce the speed of the cleansing liquid flow by the pump 8 when bubbles are detected by the sensor 9. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a cell washing apparatus.

  Conventionally, adipose tissue is decomposed by digestive enzymes to separate adipose-derived cells, and a centrifuge container containing the resulting cell suspension is subjected to centrifugal motion around an axis away from the centrifuge container. A cell separation device that separates components contained in a suspension by specific gravity is known (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. Then, the separated supernatant is discarded, the washing liquid is supplied, resuspended, and the operation of centrifuging is repeated, whereby the separated cells can be washed to reduce the concentration of the digestive enzyme solution.

International Publication No. 2005/012480 Pamphlet

  However, when supplying the washing liquid to the pellet-like cells after removing the supernatant, if bubbles are included in the supply pipe for supplying the washing liquid, the cells are damaged by the impact of the washing liquid intermittently caused by the bubbles. There is an inconvenience of receiving. In particular, if the cells are exposed to a high concentration of digestive enzymes during decomposition for a long time, the cells are damaged, so it is necessary to supply the washing solution as soon as possible to dilute it.

  The present invention has been made in view of the above-described circumstances, and prevents the cleaning liquid containing bubbles from being supplied to the cells, and quickly supplies the cleaning liquid containing no bubbles to the cells. It aims at providing the cell washing device which can do.

In order to achieve the above object, the present invention provides the following means.
The present invention includes a container that contains cells, a supply pipe that supplies a cleaning liquid into the container, a pump that sends the cleaning liquid in the supply pipe toward the container, and an intermediate position of the supply pipe above the supply pipe. And a reservoir part formed of a recess that captures bubbles contained in the cleaning liquid, and the supply pipe between the pump and the reservoir part, and detects the bubbles flowing in the supply pipe Provided is a cell washing apparatus including a sensor and a control unit that controls to reduce the flow rate of the washing liquid by the pump when bubbles are detected by the sensor.

  According to the present invention, when the cleaning liquid is sent toward the container by the operation of the pump, when the cleaning liquid does not include bubbles, the cleaning liquid does not decrease the flow rate by the pump. The digestive enzyme concentration of the cell suspension can be rapidly reduced. On the other hand, when bubbles are included in the cleaning liquid, the bubbles are detected by a sensor provided in the supply pipe, and the flow rate of the cleaning liquid by the pump is reduced by the control unit. As a result, the bubbles are easily raised by buoyancy in the reservoir portion formed of a recess provided in the middle of the supply pipe, and are more reliably captured. Therefore, it is possible to prevent the cleaning liquid containing bubbles from being supplied to the cells, and to prevent the occurrence of inconvenience that the cells are damaged by the impact of the cleaning liquid intermittently caused by the bubbles.

In the said invention, the said container is a centrifuge container, You may provide the centrifuge which carries out the centrifugal motion of this centrifuge container so that the bottom face may go to radial direction outward.
In this way, the cell suspension suspended in the centrifuge container by the washing liquid is centrifuged by the operation of the centrifuge, and the supernatant is discarded, leaving only the cells in the container. Can do. Then, by repeatedly supplying the washing liquid, centrifuging, and discarding the supernatant, the concentration of the digestive enzyme adhering to the cells can be reduced and the cells can be maintained in a healthy state.

    According to the present invention, it is possible to prevent the cleaning liquid containing bubbles from being supplied to the cells and to quickly supply the cleaning liquid containing no bubbles to the cells.

Hereinafter, a cell washing apparatus 1 according to an embodiment of the present invention will be described with reference to the drawings.
As shown in FIG. 1, the cell washing apparatus 1 according to the present embodiment has a centrifuge container 2 for storing a cell suspension, and the centrifuge container 2 has a bottom surface in the radial direction as indicated by a broken line. A centrifuge 3 that performs centrifugal movement outward, a supernatant suction tube 5 that aspirates and discards the supernatant 4 (see FIG. 2) that is placed in the centrifuge container 2 and centrifuged, and the supernatant 4 A supply pipe 7 for supplying a cleaning liquid to the cells 6 accommodated in the centrifuge container 2 after being sucked, a pump 8 for supplying the cleaning liquid in the supply pipe 7 toward the centrifuge container 2, and a supply A bubble sensor 9 for detecting a bubble A in the tube 7 (see FIG. 4), a reservoir unit 10 composed of a recess recessed upward at a midway position of the supply tube 7, and a control unit 11 for controlling the pump 8. I have.

The cell suspension is, for example, a mixture of adipose tissue collected from the human body as a living tissue and digestive enzyme, and then digesting the adipose tissue to separate adipose-derived cells. It contains the tumescent solution and digestive enzymes used in the treatment, and the fat-derived cells separated.
Adipose-derived cells include vascular endothelial cells, fibroblasts, hematopoietic stem cells, hematopoietic cells, adipose-derived stem cells, and the like. Here, the 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.

  As shown in FIG. 2, the centrifuge container 2 is formed in a tapered shape whose bottom surface tapers toward the tip, and is centrifuged by the operation of the centrifuge 3, so that the inside of the cell suspension These substances are separated in a stacked state in the depth direction of the centrifuge container 2 due to the difference in specific gravity. At this time, the cells 6 having a large specific gravity are collected at the tip along the tapered bottom surface of the centrifuge container 2, and a liquid component having a small specific gravity (supernatant 4) is separated above the cells. Yes.

  The centrifuge 3 is connected to a motor 12, an arm 13 that is rotationally driven by the motor 12 and supports the centrifuge container 2 at both ends in a swingable manner, and a supply pipe 7, and is sent via the supply pipe 7. And a rotary joint 14 that distributes the cleaning liquid to each centrifuge container 2.

  The supernatant suction tube 5 has a suction port 5 a disposed closer to the supernatant 4 than the boundary surface between the pellet-like cells 6 centrifuged in the centrifuge container 2 and the supernatant 4. Then, by sucking from the suction port 5a, the supernatant 4 can be sucked and discarded through a path (not shown) until the cells 6 and some of the supernatant 4 remain in the centrifuge container 2. Yes.

  The supply pipe 7 is at least partially made of a transparent material, and a discharge port 7 a is arranged in the cell 6 that has been centrifuged in the centrifuge container 2. By operating the pump 8, the washing liquid is discharged from the discharge port 7a of the supply pipe 7, so that the cells 6 formed in a pellet form are released and resuspended, thereby reducing the concentration of the digestive enzyme. A cell suspension is produced.

The pump 8 is a peristaltic pump, for example, and supplies the cleaning liquid at a flow rate corresponding to the rotational speed.
The bubble sensor 9 includes a light emitting portion 9a and a light receiving portion 9b that are opposed to each other with the transparent portion of the supply tube 7 interposed therebetween, and allows the bubble A to pass through the supply tube 7 in accordance with a change in the amount of light received by the light receiving portion 9b. It comes to detect.

As shown in FIG. 3, the reservoir unit 10 is disposed between the bubble sensor 9 and the centrifuge container 2. The reservoir unit 10 is provided with an air vent filter 15 at the top thereof. When bubbles A are included in the cleaning liquid that can flow in the supply pipe 7, the bubbles A that have reached the reservoir unit 10 rise in the reservoir unit 10 and accumulate in the upper part via the air vent filter 15. Are released to the outside.
The control unit 11 is connected to the bubble sensor 9 and controls the rotational speed of the pump 8 so as to reduce the flow rate of the cleaning liquid in response to the detection signal of the bubble A from the bubble sensor 9.

The operation of the cell washing apparatus 1 according to the present embodiment configured as described above will be described below.
In order to wash the cells 6 using the cell washing apparatus 1 according to the present embodiment, the cell suspension obtained by decomposing the adipose tissue is accommodated in the centrifuge container 2 and the centrifuge 3 is operated. Then, the cells 6 and the supernatant 4 are centrifuged, and the supernatant 4 is aspirated and discarded from the suction port 5a of the supernatant suction tube 5, thereby leaving the pellet-like cells 6 in the centrifuge container 2. Next, the pump 8 is operated to discharge the cleaning liquid from the discharge port 7a of the supply pipe 7 into the centrifuge container 2, and the pellet-like cells 6 group are released and resuspended.

  By repeating the centrifugation, the supernatant 4 suction, and the discharge of the washing liquid, the concentration of the digestive enzyme adhering to the cells 6 can be reduced, and the cells 6 can be washed and maintained in a healthy state.

  In this case, when the controller 11 drives the pump 8 to discharge the cleaning liquid from the discharge port 7a of the supply pipe 7, if the bubbles A are not included in the cleaning liquid, as shown in FIG. The pump 8 is rotated at a high speed to quickly supply the cleaning liquid. On the other hand, when the bubble sensor 9 detects the bubble A in the cleaning liquid, the control unit 11 decreases the rotational speed of the pump 8 as shown in FIG. 4 to decrease the flow rate of the cleaning liquid.

  When the cleaning liquid contains bubbles A, if the cleaning liquid is supplied at a high speed, the bubbles A cannot reach the reservoir unit 10 even if they reach the reservoir unit 10 and ride on the flow of the cleaning liquid to the tip of the supply pipe 7. It is sent and discharged from the discharge port 7a. When the bubbles A are discharged from the discharge port 7a, the discharge of the cleaning liquid is interrupted, so that the impact is applied to the cells 6 and the cells 6 are damaged.

  On the other hand, according to the present embodiment, when the bubble sensor 9 detects the bubble A, the flow rate of the cleaning liquid is reduced, so that the bubble A moves away from the flow of the cleaning liquid when it reaches the reservoir unit 10. The inside of the reservoir unit 10 is raised and discharged from the air vent filter 15 to the outside. Accordingly, there is an advantage that the bubbles A are prevented from being discharged from the discharge port 7a of the supply pipe 7, and the cells 6 are not damaged.

  As described above, according to the cell cleaning device 1 according to the present embodiment, when the bubbles A are included in the cleaning liquid, the flow rate is decreased so that the bubbles A are not discharged from the discharge port 7a. When A is not included, the flow rate can be increased to supply the washing solution quickly, and the concentration of the digestive enzyme attached to the cells 6 can be rapidly reduced. That is, both damage caused by intermittent discharge of the cleaning liquid and damage caused by digestive enzymes can be prevented from being applied to the cell 6, and the cell 6 can be maintained in a healthy state.

  In the present embodiment, the case of washing a cell suspension containing fat-derived cells separated from adipose tissue has been described. However, the present invention is not limited to this and is used for washing other cells 6. May be.

It is a whole lineblock diagram showing the cell washing device concerning one embodiment of the present invention. It is a longitudinal cross-sectional view which shows the centrifuge container with which the cell washing apparatus of FIG. 1 was equipped. It is a figure explaining the supply operation | movement of the washing | cleaning liquid which does not contain a bubble in the cell washing | cleaning apparatus of FIG. It is a figure explaining supply operation | movement of the washing | cleaning liquid containing a bubble in the cell washing | cleaning apparatus of FIG.

Explanation of symbols

A Bubble 1 Cell washing device 2 Centrifugal container (container)
3 Centrifuge 6 Cell 7 Supply Pipe 8 Pump 9 Bubble Sensor (Sensor)
10 Reservoir unit 11 Control unit

Claims (2)

A container for containing cells;
A supply pipe for supplying a cleaning liquid into the container;
A pump for sending the cleaning liquid in the supply pipe toward the container;
A reservoir part that is provided at an intermediate position of the supply pipe so as to extend above the supply pipe, and is formed of a recess for capturing bubbles contained in the cleaning liquid;
A sensor that is disposed in the supply pipe between the pump and the reservoir and detects bubbles flowing in the supply pipe;
A cell cleaning apparatus comprising: a control unit that controls to decrease a flow rate of the cleaning liquid by the pump when bubbles are detected by the sensor. The container is a centrifuge container;
The cell washing apparatus according to claim 1, further comprising a centrifuge that centrifuges the centrifuge container so that a bottom surface thereof faces radially outward.

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