JP2009189280A - Centrifugal separation container and centrifugal separation apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a centrifugal separation container and a centrifugal separation apparatus that easily and surely take out a substance centrifuged from a liquid and in the operation, avoid substance contaminations and maintain soundness of substance. <P>SOLUTION: The centrifugal separation container 1 includes a container body 3 that holds a liquid A to be separated and has a bottom part 3b to be rotated outward in the radial direction and is equipped with an output port 4, in the vicinity of the bottom part 3b of the container body 3, which is passed through a container wall and takes out a substance in the inside. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a centrifuge container and a centrifuge.

Conventionally, a centrifuge device that separates components contained in a liquid by specific gravity by rotating a centrifuge container containing a liquid to be separated around an axis away from the centrifuge container 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.

WO05 / 012480 pamphlet

  However, in order to take out the components separated near the closed end after being separated into the centrifuge container from the centrifuge container, the needle of the syringe is inserted and sucked from the end opening opposite to the closed end. It is difficult to insert the needle securely up to the closed end, and there is a disadvantage that the bottom of the centrifuge container is damaged at the tip of the needle.

  In particular, when centrifuging cells from a cell suspension, it is not preferable to insert a cylinder portion that touches the operator's hand into the centrifuge container in order to avoid contamination to the cells as much as possible. Therefore, it is necessary to make the needle of the syringe extremely long and insert only the needle, and the insertion operation is further difficult. Further, when the separated cells are aspirated by an elongated needle, the cells may be damaged by shearing during the aspiration, and there is a disadvantage that the soundness of the cells is lowered.

  The present invention has been made in view of the above-described circumstances, and can easily and reliably take out a substance centrifuged from a liquid. In this case, contamination of the substance can be avoided, and An object of the present invention is to provide a centrifuge container and a centrifuge that can maintain soundness.

In order to achieve the above object, the present invention provides the following means.
The present invention includes a container main body that contains a liquid to be separated and is rotated with its bottom portion directed radially outward, and in the vicinity of the bottom portion of the container main body, takes out the substance inside through the container wall. A centrifuge container provided with a take-out port is provided.

  According to the present invention, the liquid to be separated is accommodated in the container main body, and the bottom part is rotated outward in the radial direction, whereby substances having large specific gravity are separated in order from the vicinity of the bottom part. And the substance inside a container main body can be easily taken out by penetrating a container wall with the taking-out port provided in the bottom part vicinity. There is no need to insert an elongated syringe needle from the end opposite to the bottom to suck the substance, and it can be taken out through a short path that only penetrates the container wall, maintaining the soundness of the substance. it can. Moreover, since it is not necessary to insert a syringe etc. inside a container main body, generation | occurrence | production of contamination can be prevented.

In the above invention, the bottom portion of the container main body may be formed in a tapered inner surface shape that tapers toward the tip, and the take-out port may be provided in communication with the most distal end portion of the bottom portion.
By doing in this way, the substance with large specific gravity collected along the taper inner surface by centrifuging can be taken out without leaving from the taking-out port which penetrates the most advanced part.

Moreover, in the said invention, it is good also as the suction device which suck | inhales an internal substance being connected to the said extraction opening so that isolation | separation is possible.
In this way, after the centrifugal separation is completed, the suction device is operated to suck the substance inside, and then the suction device is removed from the take-out port, and the taken-out substance is transported and supplied to the necessary site. can do.

Moreover, in the said invention, the said container main body and the said suction device are good also as being integrally shape | molded on both sides of the cutting part which comprises the discharge outlet of a suction device by cutting | disconnection.
By doing in this way, the centrifuge container which connected the container main body and the suction instrument aseptically by integral molding can be comprised. Then, after sucking the substance centrifuged in the container body by the operation of the suction instrument, the suction instrument is separated from the container body at the cutting portion, thereby forming a discharge port in the suction instrument, and the sucked substance is removed. It becomes possible to discharge from the discharge port.

Moreover, in the said invention, the said suction device is good also as consisting of a dropper or a syringe.
By doing in this way, the substance isolate | separated in the container main body inside a dropper or a syringe can be attracted | sucked easily, and the attracted | sucked substance can be discharged easily.

  The present invention also provides a centrifuge for rotating any one of the above centrifuge containers to separate the liquid components in the container body by specific gravity, and supporting the centrifuge container in a removable manner. A container support portion, an arm portion that supports the container support portion so as to be swingable about a swing axis perpendicular to the longitudinal axis, and a rotation that is disposed at a position spaced apart from the swing axis. A centrifuge provided with a driving device that rotates around an axis, and a container holding unit that includes a body holding unit that holds the container body, and a suction device holding unit that holds a suction device connected to the container body I will provide a.

  According to the present invention, when the centrifuge container is inserted into and supported by the cup-shaped container support part, the container main body is held by the main body holding part, and the suction instrument is held by the suction instrument holding part. In this state, when the driving device is operated, the container support portion that is swingably supported around the swing axis is swung by the arm portion that is rotated around the rotation axis, and the bottom portion of the container body is moved in the radial direction. A substance having a large specific gravity is centrifuged toward the bottom and directed outward. Thereafter, the drive device is stopped, the rotation of the arm portion is stopped, and the suction device is operated, so that the substance centrifuged at the bottom of the container body can be easily sucked without remaining in the suction device. .

In the said invention, the said suction device holding | maintenance part is good also as arrange | positioning with respect to the said main body holding | maintenance part at the rotation direction back side of the said arm part by the said drive device.
By doing in this way, even if the arm part and the container support part and the centrifuge container supported by the arm part are rotated at a high speed by the operation of the driving device, the air flow generated by the suction device connected to the container body Can be suppressed, and the occurrence of vibration and wind noise can be reduced.

Moreover, in the said invention, it is good also as an observation window which makes the said suction instrument holding | maintenance part visible the side surface of a suction instrument.
In this way, when the substance inside the container body is sucked by the suction device, the side surface of the suction device can be visually confirmed through the observation window, and the suction device can be made of a translucent or transparent material. For example, the presence or absence of an internal substance can be easily confirmed.

  According to the present invention, a substance centrifuged from a liquid can be easily and reliably taken out, and at that time, the contamination of the substance can be avoided and the soundness of the substance can be maintained. Play.

A centrifuge container 1 and a centrifuge device 2 according to an embodiment of the present invention will be described below with reference to FIGS.
As shown in FIG. 1, the centrifuge container 1 according to this embodiment is a liquid A to be separated (see FIG. 5), for example, cells and digestive enzymes isolated by digesting adipose tissue with digestive enzymes. And a syringe (aspirator) 5 connected to an outlet 4 provided in the container body 3.

The container body 3 is formed in a substantially cylindrical shape, has an upper opening 3a at one end, and is closed at the other end. The closed end (bottom part) 3b is formed in a tapered inner surface shape that tapers gradually toward the tip, and the outlet 4 is opened at the narrowest part of the tip.
A cap 6 is detachably placed on the upper opening 3a, and a supply pipe 7 for supplying the cell suspension A and a suction pipe 8 for sucking the liquid inside are inserted and arranged in the container body 3. It is like that.

The syringe 5 is attached to the take-out port 4 through a joint 9 so that the discharge port 10 can be attached and detached.
The syringe 5 includes a cylinder 5a and a piston 5b made of a transparent or translucent material, and when performing a centrifugal separation operation, the piston 5b is arranged at a position where the piston 5b is pushed most into the cylinder 5a. Yes. Further, when performing the centrifugal separation work, a fixing member 11 for restraining the movement of the piston 5b with respect to the cylinder 5a is attached.

  As shown in FIGS. 2 to 4, the centrifugal separator 2 according to the present embodiment includes an arm portion 13 fixed to a rotary shaft 12 along a vertical direction that is rotationally driven by a driving device such as a motor (not shown). The arm 13 is provided at both ends with a container support 15 that is rotatably supported around a swing shaft 14 that is disposed substantially horizontally.

  The container support portion 15 is formed in a cup shape that opens upward in a stationary state, and a main body holding portion 16 that holds the container main body 3 removably from above, and a syringe 5 connected to the container main body 3 upward. And a syringe holding part (suction device holding part) 17 that is detachably held. The syringe holding part 17 is arrange | positioned in the position twisted to the rotation direction back side of the arm part 13 around the rotating shaft 12 with respect to the main body holding part 16, as FIG. 3 shows.

  In addition, an observation window 18 that exposes the side surface of the cylinder 5a that is held is provided at an intermediate position of the syringe holding portion 17. Since the cylinder 5a is made of a transparent or translucent material, when the side surface of the cylinder 5a is observed through the observation window 18, the presence or absence of the substance sucked into the cylinder 5a can be easily confirmed. .

The operation of the centrifuge container 1 and the centrifuge device 2 according to this embodiment configured as described above will be described below.
In order to obtain a desired cell, for example, a stem cell B, by centrifuging the cell suspension A using the centrifuge container 1 and the centrifuge 2 according to the present embodiment, first, as shown in FIG. Then, the centrifuge container 1 in which the cell suspension A is accommodated in the container body 3 through the supply pipe 7 is inserted into the container support 15 of the centrifuge 2 and supported as shown in FIG. .

  Since the container support portion 15 is provided with a cup-shaped main body holding portion 16 that holds the container main body 3 and a syringe holding portion 17 that holds the syringe 5, the centrifuge container 1 is placed above the container support portion 15. The container main body 3 is stably held by the main body holding portion 16 and the syringe 5 is stably held by the syringe holding portion 17. In this state, the drive device is operated and, as shown in FIG. 3, the arm 13 is rotated about the rotating shaft 12 to swing the container support 15 around the swinging shaft 14 by centrifugal force. Then, the container body 3 is rotated at a predetermined rotation speed and time so that the bottom 3b of the container body 3 is directed outward in the radial direction.

  Since the bottom 3b of the container body 3 is formed in a tapered inner surface, stem cells B having a large specific gravity contained in the cell suspension A are collected along the tapered inner surface at the tip of the bottom 3b. Thereby, as shown in FIG. 6, stem cells B having a large specific gravity contained in the cell suspension A sink to the tip of the bottom 3 b of the container body 3 and are separated from the remaining supernatant C.

  At this time, according to the centrifugal separator 2 according to the present embodiment, since the syringe 5 is attached to the container body 3, when the arm portion 13 is rotated, the container body 3 and the syringe 5 are rotated together. . In this case, since the syringe 5 is disposed rearward in the rotation direction with respect to the container main body 3, even if the arm portion 13 is rotated at a high speed, the disturbance of the airflow by the syringe 5 protruding from the container main body 3 is alleviated. The generation of large vibrations and wind noises is prevented.

  In this state, as shown in FIG. 7, the supernatant C in the container body 3 is sucked through the suction tube 8, and the separated stem cells B and a small amount of the supernatant C remain to be supplied. A cleaning solution such as Ringer's solution is supplied from the tube 7 and stirred again. As a result, the state of FIG. 5 is obtained again, so that the stem cell B is washed by repeating centrifugation, suction, and re-stir several times, and the concentration of the digestive enzyme contained in the cell suspension A is sufficiently high. Can be reduced.

After a plurality of operations are repeated, when the state of FIG. 7 is obtained by suction of the supernatant C, the fixing member 11 is removed from the syringe 5 and, as shown in FIG. 8, the piston 5b is attached to the cylinder 5a. By reciprocating, a small amount of cell suspension A ′ in which a small amount of supernatant C and stem cell B are suspended is generated.
At this time, in the centrifugal separator 2 according to the present embodiment, since the observation window 18 is provided in the syringe holding part 17, the state of the cell suspension A ′ sucked into the syringe 5 is changed to the observation window 18. Can be easily confirmed.

In this state, as shown in FIG. 9, by moving the piston 5 b in the direction in which the piston 5 b is pulled out with respect to the cylinder 5 a, a small amount of the cell suspension A ′ stored in the container body 3 is removed from the syringe 5. Can be sucked in.
With all the cell suspension A ′ sucked into the syringe 5, as shown in FIG. 10, by removing the syringe 5 from the joint 9, it can be directly transported to another place and discharged. It becomes.

  Thus, according to the centrifuge container 1 according to the present embodiment, since the outlet 4 is provided through the container wall at the tip of the container body 3, and the syringe 5 is connected to the outlet 4, Stem cells B centrifuged in the container body 3 can be taken out from the outlet 4 into the syringe 5 through a short path. That is, since the path is short, the shearing force received by the stem cell B in the narrow path can be reduced, and the stem cell B can be sucked into the syringe 5 while maintaining a healthy state.

  Further, since the centrifuged stem cell B is taken out from the outlet 4 provided at the tip of the container body 3, it is not necessary to insert and remove the syringe 5 from the upper opening 3a of the container body 3. That is, when the syringe 5 is inserted from the upper opening 3a, the distal end of the needle must be accurately placed at the lowest position on the bottom surface, and the centrifuged stem cells B cannot be sucked without remaining. According to the embodiment, it is possible to suck out and extract all stem cells without performing such difficult and troublesome work.

  Further, when a suction device such as a syringe is inserted from the upper opening 3a of the container body 3 and the internal stem cells B are sucked, the possibility that dust, bacteria, etc. attached to the syringe will enter the container body 3 is increased. It is not preferable. According to this embodiment, it is not necessary to insert a suction device from the upper opening 3a, and contamination can be prevented from occurring.

  In the present embodiment, the syringe 5 is detachably connected to the take-out port 4 provided in the bottom 3b of the container body 3 via the joint 9, but this is shown in FIG. As described above, the one in which the outlet 4 of the container body 3 and the cylinder 5a of the syringe 5 are connected by an elongated connecting pipe (cutting part) 20 is integrally formed by, for example, resin molding, and the piston 5b is formed in the cylinder 5a. It may be inserted.

  In this way, after the cell suspension A ′ containing the stem cell B centrifuged inside the container body 3 is sucked into the syringe 5, the middle position of the elongated connecting tube 20 is cut, thereby the syringe 5. Can be easily separated. Thereby, the centrifuge container 1 can be made into a disposable product. In this case, the cut portion of the connection tube 20 can be used as a discharge port for discharging the cell suspension A ′ accommodated in the syringe 5. Moreover, it can be made to function similarly as a dropper instead of the syringe 5. Moreover, although cell suspension A was illustrated as a liquid which should be isolate | separated, it is not limited to this.

It is a longitudinal section showing a centrifuge container concerning one embodiment of the present invention. It is a front view which shows a part of centrifuge which concerns on one Embodiment of this invention holding the centrifuge container of FIG. It is a top view of the centrifuge of FIG. FIG. 3 is a perspective view of the centrifugal separator in FIG. 2. It is a longitudinal cross-sectional view which shows the state which supplied the cell suspension to the centrifuge container of FIG. It is a longitudinal cross-sectional view which shows the state after the centrifugation by the centrifuge container of FIG. It is a longitudinal cross-sectional view which shows the state which attracted | sucked the supernatant in the centrifuge container of FIG. It is a longitudinal cross-sectional view which shows the state which stirred the small amount supernatant and stem cell in the centrifuge container of FIG. 7 by the reciprocating operation of the syringe. It is a longitudinal cross-sectional view which shows the state which attracted | sucked the cell suspension in the centrifuge container of FIG. 8 in the syringe. FIG. 10 is a longitudinal sectional view showing a state where a syringe that sucked the cell suspension in the centrifuge container of FIG. 9 is removed. It is a longitudinal cross-sectional view which shows the modification of the centrifuge container of FIG.

Explanation of symbols

A, A 'Cell suspension (liquid)
B Stem cell (substance)
DESCRIPTION OF SYMBOLS 1 Centrifugal container 2 Centrifugal apparatus 3 Container main body 3b Bottom part 4 Outlet 5 Syringe (suction device)
DESCRIPTION OF SYMBOLS 10 Discharge port 12 Rotation axis 13 Arm part 14 Oscillation axis 15 Container support part 16 Main body holding part 17 Syringe holding part (suction instrument holding part)
18 Observation window 20 Connecting pipe (cutting part)

Claims (8)

A container body that contains the liquid to be separated and can be rotated with the bottom facing radially outward;
A centrifuge container in which a take-out port is provided in the vicinity of the bottom of the container main body for taking out an internal substance through the container wall. The bottom of the container body is formed in a tapered inner surface that tapers toward the tip,
The centrifuge container according to claim 1, wherein the take-out port is provided in communication with the most distal end portion of the bottom portion.   The centrifuge container according to claim 1 or 2, wherein a suction device for sucking an internal substance is detachably connected to the extraction port.   The centrifuge container according to claim 3, wherein the container body and the suction device are integrally formed with a cutting portion constituting a discharge port of the suction device being cut.   The centrifuge container according to claim 3 or 4, wherein the suction device comprises a dropper or a syringe. A centrifuge for rotating the centrifuge container according to any one of claims 3 to 5 to separate liquid components in the container body by specific gravity,
A cup-shaped container support part for removably supporting the centrifuge container;
An arm portion that supports the container support portion so as to be swingable about a swing axis perpendicular to the longitudinal axis;
A drive device that rotationally drives the arm portion around a rotation axis disposed at a position spaced apart from the swing axis;
A centrifugal separator comprising a main body holding portion for holding the container main body and a suction device holding portion for holding a suction device connected to the container main body on the container support portion.   The centrifuge according to claim 6, wherein the suction device holding part is disposed on the rear side in the rotation direction of the arm part by the driving device with respect to the main body holding part.   The centrifuge according to claim 6 or 7, wherein an observation window that makes the side surface of the suction device visible is provided in the suction device holding portion.

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