The present invention relates to a device for producing a three dimensional structural cell and more particularly to a device for producing a three-dimensional structural cell which produces a three-dimensional structural body by a plurality of cells by sticking a needle-shaped body provided on a support into a cell mass.
With the purpose of use for medical cell transplant for reproduction of organs or internal organs and for other tests, production of a three-dimensional structural body of cells has been practiced by combining a plurality of cell masses (spheroids) in a three-dimensional manner.
In order to produce such three-dimensional structural bodies of cells, use of a support body provided with a plurality of needle-shaped bodies is known, and the needle-shaped bodies are stuck into the plurality of cell masses so as to bring the cell masses close to each other, whereby the three-dimensional structural bodies of the cells are obtained.
In the conventional arts, in order to stick the needle-shaped body of the support body into the cell masses, a-manual work was performed by using a pipette having suctioned the cell masses, a robot arm, and tweezers (Patent Literature 1).
Patent Literature 1: Japanese Patent No. 4517125
However, the cell masses have a diameter of approximately several hundreds μm, and there was a problem that the work to stick the needle-shaped body having a diameter of several tens μm into them was very difficult.
In view of the above problem, the present invention provides a device for producing a three-dimensional structural cell which can obtain the three-dimensional structural body of a cell by automatically sticking the needle-shaped body of the support body into the cell masses.
That is, a device for producing a three-dimensional structural cell according to the present invention of claim (1) is provided with an accommodating plate in which a large number of accommodating recess portions, each accommodating one cell mass, are formed, a support body provided with a plurality of needle-shaped bodies sticking and penetrating the cell mass, a suction nozzle for adsorbing/holding the cell mass by being connected to negative pressure generating means, moving means for moving and elevating the suction nozzle between the accommodating plate and the support body, and control means for controlling a suctioning operation of the suction nozzle and an operation of the moving means, in which
the suction nozzle is provided with a tubular adsorbing portion for adsorbing one cell mass at a tip end thereof, an inner diameter of the adsorbing portion is formed smaller than an outer diameter of the cell mass and larger than an outer peripheral diameter of the needle-shaped body of the support body;
the control means is configured such that, when the cell mass is adsorbed/held by the adsorbing portion of the suction nozzle in the accommodating recess portion of the accommodating plate, the suction nozzle is moved to above the required needle-shaped body in the support body by the moving means; and
the suction nozzle is further moved in an axial direction of the needle-shaped body, the needle-shaped body is stuck into the cell mass, and the needle-shaped body penetrating the cell mass is inserted into the adsorbing portion.
According to the above invention, since the suction nozzle is moved by the moving means so that the cell mass in the accommodating plate is adsorbed/held and is stuck by the needle-shaped body of the support body, production of a three-dimensional structural body by the cell mass can be performed automatically.
At that time, since the inner diameter of the adsorbing portion of the adsorption nozzle is formed smaller than the outer diameter of the cell mass and larger than the outer peripheral diameter of the needle-shaped body and the moving means moves the suction nozzle having adsorbed/held the cell mass in the axial direction of the needle-shaped body, the cell mass can be reliably stuck by the needle-shaped body.
An illustrated embodiment will be described below, in which
In this device 2 for producing a three-dimensional structural body, by using a suction nozzle 19 which will be described later in detail, a cell mass 1 is taken out of an accommodating plate 3 illustrated in
Moreover, in the present embodiment, it is possible to produce the three-dimensional structural body 1A by mixing partially different kinds of different-kind cell masses 1b, and thus, the accommodating plate 3 accommodating the cell mass 1 and a different-kind accommodating plate 3b for accommodating the different-kind cell mass 1b can be prepared.
The support body 4 is composed of a substantially regular square plate-shaped member 5 and a plurality of the needle-shaped bodies 6 installed upright on a surface of the plate-shaped member 5, and in the present embodiment, the needle-shaped bodies 6 are provided in the number of 5 pieces in a vertical direction and 5 pieces in a lateral direction, totaling 4-25 pieces, on the surface of the plate-shaped member 5. The number of the needle-shaped bodies 6 is not limited to this.
This support body 4 is accommodated in an accommodating vessel 7 illustrated in
The needle-shaped body 6 is made of metal and has a length with which it can stick a plurality of (4 pieces in this embodiment) cell masses 1, respectively, and an interval between the adjacent needle-shaped bodies 6 is set at such an interval that the stuck cell masses 1 are brought into close contact with each other, that is, an interval of approximately one cell mass 1.
By sticking each of the needle-shaped bodies 6 into the plurality of cell masses 1 and by stacking the cell masses 1 from a root side to a tip end side of the needle-shaped body 6, the three-dimensional structural body 1A of a cell as illustrated in
Though the needle-shaped body 6 is installed substantially perpendicularly to the plate-shaped member 5, the largely inclined needle-shaped body 6 indicated by A in
The device 2 for producing a three-dimensional structural body is provided with a clean bench 11 having an inside kept in an aseptic state, a supply magazine 12 for accommodating an accommodating plate 3 which has accommodated the cell mass 1, a recovery magazine 13 for recovering the accommodating plate 3 from which the cell mass 1 has been taken out, first support means 14 for supporting the accommodating plate 3 arranged below the recovery magazine 13, second support means 15 for supporting a different-kind accommodating plate 3b accommodating the different-kind cell mass 1b, third support means 16 arranged adjacently to the support body 4 and supporting the accommodating plate 3 and the different-kind accommodating plate 3b, transfer means 17 for transferring the accommodating plate 3 and the different-kind accommodating plate 3b, a placing base 18 on which the accommodating vessel 7 accommodating the support body 4 is placed, a suction nozzle 19 for adsorbing/holding the cell mass 1, and moving means 20 for moving and elevating the suction nozzle 19.
The device 2 for producing a three-dimensional structural body having the aforementioned configuration is controlled by control means 21 provided adjacently to the clean bench 11, and this control means 21 is capable of various settings by a terminal such as a personal computer 21a or the like.
The clean bench 11 is capable of being opened/closed by an opening/closing door, not shown, and of carrying′ in/out the supply magazine 12, the recovery magazine 13, and the accommodating vessel 7 accommodating the support body 4.
On an upper part of the clean bench 11, aseptic air supply means, not shown, for supplying an aseptic air is provided so that, in a state in which the opening/closing door is closed, a one-way flow of the aseptic air from an upper side to a lower side is formed inside the clean bench 11.
The supply magazine 12 and the recovery magazine 13 are capable of holding the plurality of accommodating plates 3 in a stacked state, respectively, and the second support means 15 is provided at a position adjacent to a side of the recovery magazine 13.
The first and third support means 14 and 16 are arranged at the same height and constituted by four support members supporting four corners of the accommodating plate 3 from below, and the transfer means 17 is configured to place the accommodating plate 3 from above on the first and third support means 14 and 16.
On the other hand, the second support means 15 is provided slightly above the first and third support means 14 and 16 and is provided with an engaging claw 15a capable of advancing/retracting for supporting the four corners of the different-kind accommodating plate 3b from below.
The second support means 15 is configured to support the different-kind accommodating plate 3b from below by protrusion of the engaging claw 15a, and when the different-kind accommodating plate 3b is transferred from the second support means 15 to the transfer means 17 or when the different-kind accommodating plate 3b is transferred from the transfer means 17 to the second support means 15, the engaging claw 15a is retracted so as to allow vertical movement of the different-kind accommodating plate 3b.
The transfer means 17 is composed of a slide mechanism 22 provided linearly below the supply magazine 12, the recovery magazine 13, and the first to third support means 14 to 16 and an elevating table 24 provided on a movable portion 23 of the slide mechanism 22.
The elevating table 24 lowers when the first to third support means 14 to 16 support the accommodating plate 3 or the different-kind accommodating plate 3b and is located above the first and third support means 14 and 16 and below the second support means 15 when the accommodating plate 3 or the different-kind accommodating plate 3b are transferred and moves the accommodating plate 3 or the different-kind accommodating plate 3b by an operation of the slide mechanism 22 in that state.
On the placing base 18, the accommodating vessel 7 for accommodating the support body 4 is positioned and placed, and a ring illumination 25 for illuminating the accommodating vessel 7 is provided through turning means 26 for turning/moving the ring illumination 25 so that the ring illumination 25 is turned/moved between above the accommodating vessel 7 and a position retreated therefrom.
As illustrated in
The suction nozzle 19 is provided with a tubular adsorbing portion 19a for adsorbing one cell mass at its tip end and a body portion 19b held by the moving means 20 and is attached to the moving means 20 by fixing means 20a, capable of replacement.
The adsorbing portion 19a has its inner diameter smaller than an outer diameter of the cell mass 1 and larger than an outer peripheral diameter of the needle-shaped body 6 of the support body 4. By setting such dimensions, as illustrated in
The moving means 20 is composed of an X-Y unit 34 as a horizontal moving mechanism provided at a position adjacent to the transfer means 17 as illustrated in
The X-Y unit 34 is composed of a Y-direction mechanism 34a for moving the movable support portion 35 in a direction orthogonal to a transfer direction of the transfer means 17 and an X-direction mechanism 34b for moving the Y-direction mechanism 34a in a direction orthogonal to the moving direction.
On a movable portion of the elevating means 36, the suction nozzle 19 is fixed through the fixing means 20a, and by moving the movable support portion 35 in a horizontal direction by the X-Y unit 34, the suction nozzle 19 can be moved between the accommodating plate 3 or the different-kind accommodating plate 3a supported by the third support means 16 and the accommodating vessel 7 placed on the placing base 18, and also by vertically elevating the suction nozzle 19 by the elevating means 36, the cell mass 1 or different-kind cell mass 1b in the accommodating recess portion 3a of the accommodating plate 3 or the different-kind accommodating plate 3b can be adsorbed or held, or the held cell mass 1 or different-kind cell mass 1b can be stuck by the needle-shaped body 6 of the support body 4.
Above the third support means 16, a panel illumination 37 is provided and performs plane light emitting with substantially the same area as that of the accommodating plate 3 or the different-kind accommodating plate 3b supported by the third support means 16 and emits light onto the entirety of the accommodating plate 3 or the different-kind accommodating plate 3b.
Moreover, the panel illumination 37 is reciprocally moved by a slide mechanism 37a from above the third support means 16 to a side in the transfer direction of the transfer means 17 and is capable of retreating so as not to interfere with the lowering of the suction nozzle 19 to the accommodating plate 3 or the different-kind accommodating plate 3b supported by the third support means 16.
On the upper part of the movable support portion 35, a mounting member 35a which is long downward is provided side by side with the elevating means 36, and a lower camera 38 is mounted on a lower part of the mounting member 35a, and an upper camera 39 on the upper part, respectively.
The lower camera 38 is arranged so as to be located immediately below the suction nozzle 19, and the accommodating plate 3 or the different-kind accommodating plate 3b supported by the third support means 16 and the panel illumination 37 are located between the lower camera 38 and the suction nozzle 19.
The lower camera 38 photographs the transparent accommodating plate 3 or different-kind accommodating plate 3b from below. Its photographing range is, as illustrated in
An image photographed by the lower camera 38 is transmitted to the control means 21, and the control means 21 recognizes the position in the accommodating recess portion 3a of the cell mass 1 accommodated in this accommodating recess portion 3a and also recognizes an outer diameter and a shape of the cell mass 1.
Moreover, in a state in which the accommodating plate 3 or the panel illumination 37 is not located between them, the tip end of the adsorbing portion 19a attached to the suction nozzle 19 as illustrated in
The upper camera 39 is located above the support body 4 accommodated in the accommodating vessel 7 positioned on the placing base 18 by an operation of the X-Y unit 34 of the moving means 20 and is configured to photograph all the needle-shaped bodies 6 as illustrated in
An image photographed by the upper camera 39 is transmitted to the control means 21, and the control means 21 recognizes a tip-end position of the needle-shaped body 6 provided on this support body 4.
An operation of the device 2 for producing a three-dimensional structural body having the aforementioned configuration will be described below.
First, a worker sets the supply magazine 12 accommodating the accommodating plate 3 accommodating the cell mass 1 and the empty recovery magazine 13 and also places the different-kind accommodating plate 3b accommodating the different-kind cell mass 1b on the second support means 15. Moreover, the worker positions the accommodating vessel 7 accommodating the support body 4 on the placing base 18 and attaches the new adsorbing portion 19a to the suction nozzle 19.
Moreover, by using the personal computer 21a connected to the control means 21 in advance, a structure of the three-dimensional structural body 1A of a cell to be produced is set, and arrangement of the cell mass 1 and the different-kind cell mass 1b is set as illustrated in
First, the control means 21 measures a position of the tip end of the adsorbing portion 19a of the attached suction nozzle 19. Specifically, in a state in which the panel illumination 37 is retracted, the adsorbing portion 19a of the suction nozzle 19 is photographed by the lower camera 38, and an image illustrated in
The control means 21 measures how much a center of the adsorbing portion 19a is shifted with respect to the center of the visual field of the lower camera 38 through image processing and stores the shift amount. This shift amount is added when the suction nozzle 19 is positioned at positions of the cell mass 1 or the different-kind cell mass 1b and the needle-shaped body 6 by the operation of the X-Y unit 34 of the moving means 20.
Subsequently, the elevating table 24 of the transfer means 17 is elevated, the single accommodating plate 3 is taken out of the supply magazine 12, and the accommodating plate 3 is transferred to the third support means 16 as it is.
When the accommodating plate 3 is supported by the third support means 16 as above, the panel illumination 37 is moved to above the accommodating plate 3, the suction nozzle 19 and the lower camera 38 are moved in the horizontal direction by the operation of the X-Y unit 34 of the moving means 20, and they are located above and below the accommodating recess portion 3a accommodating the cell mass 1 to be taken out.
At this time, the X-Y unit 34 moves the lower camera 38 in accordance with the position of each of the accommodating recess portions 3a of the accommodating plate 3 stored in advance and positions the entire accommodating recess portion 3a within the visual field of the lower camera 38.
When the lower camera 38 is to photograph the accommodating recess portion 3a, the panel illumination 37 emits light from above the accommodating plate 3 and can clearly recognize the accommodated cell mass 1 from below the transparent accommodating plate 3, and the control means 21 recognizes the position of the photographed cell mass 1 in the accommodating recess portion 3a and measures the size (outer diameter dimension) and the shape.
Each of
The control means 21 determines that such cell masses 1 are not to be used for the three-dimensional structural body 1A of a cell and does not perform an adsorbing/holding operation by the suction nozzle 19.
On the other hand,
Since the bottom surface of the accommodating recess portion 3a of the accommodating plate 3 has a semi-spherical shape, if the cell mass 1 is located at the center as in
The control means 21 grasps a depth corresponding to a position on the bottom surface of the accommodating recess portion 3a. Moreover, since the cell mass 1 has a substantially spherical shape, its height can be estimated from the outer-diameter dimension (diameter). The control means 21 sets a lowering amount of the suction nozzle 19 when the cell mass 1 is adsorbed/held, by recognizing the position in the accommodating recess portion 3a and by measuring the outer-diameter dimension of the cell mass 1.
After the cell mass 1 in the accommodating portion 3a is photographed by the lower camera 38 as above, the control means 21 retracts the panel illumination 37 from above the accommodating plate 3, moves the suction nozzle 19 in the horizontal XY direction by operating the X-Y unit 34 of the moving means 20, and positions it to a position of the recognized cell mass 1 and moreover, lowers the suction nozzle 19 by the elevating means 36 in accordance with the set lowering amount and brings the adsorbing portion 19a closer to the cell mass 1 so as to have the adsorbing portion 19a absorb the cell mass 1.
Since the outer diameter of the cell mass 1 is larger than the inner diameter of the adsorbing portion 19a, the cell mass 1 can be adsorbed/held at the tip end without being suctioned into the adsorbing portion 19a.
After the cell mass 1 is adsorbed/held by the suction nozzle 19 as above, the suction nozzle 19 is raised by the elevating means 36, and the panel illumination 37 is located above the accommodating plate 3 again.
The lower camera 38 photographs the accommodating recess portion 3a this time again and confirms that the cell mass 1 has been removed. If the cell mass 1 remains, a process of adsorbing/holding the cell mass 1 by the suction nozzle 19 is retried again.
When the suction nozzle 19 adsorbs and holds the cell mass 1 as above, the control means 21 operates the X-Y unit 34 of the moving means 20 so as to move the upper camera 39 to above the support body 4 and recognizes the tip end position of the needle-shaped body 6 for sticking the cell mass 1 this time.
At this time, the ring illumination 25 is located above the accommodating vessel 7 and illuminates the support body 4 and when the tip end position of the needle-shaped body 6 is recognized by the upper camera 39, the ring illumination 25 is retreated by the turning means 26.
If the tip end position of the needle-shaped body 6 is recognized, the control means 21 operates the X-Y unit 34 so as to move the adsorbing portion 19a of the suction nozzle 19 adsorbing/holding the cell mass 1 to above the tip end position of the needle-shaped body 6 recognized as above on the support body 4 and moreover, lowers the suction nozzle 19 so as to stick the cell mass 1 by the tip end of the needle-shaped body 6 as illustrated in
From this state, when the elevating means 36 moves the suction nozzle 19 downward as illustrated in
Moreover, the lowering amount of the suction nozzle 19 is set in advance but is configured to be modified on the basis of the outer diameter dimension of the cell mass 1 previously recognized by the lower camera 38.
Here, the inner diameter of the adsorbing portion 19a of the adsorption nozzle 19 in this embodiment is set smaller than the cell mass 1 and larger than the outer peripheral diameter (thickness) of the needle-shaped body 6, and the moving means 20 moves the suction nozzle 19 adsorbing/holding the cell mass 1 in an axial direction of the needle-shaped body 6, and thus, the cell mass 1 can be reliably stuck by the needle-shaped body 6.
On the other hand, as the result of recognition of the tip end position of the needle-shaped body 6, if the tip end position of the needle-shaped body 6 to stick the cell mass 1 is out of the specified range, the control means 21 has the cell mass 1 stuck at the tip end position of the needle-shaped body 6 out of the range as illustrated in
An operation when the different-kind cell mass 1b is transferred onto the support body 4 will be described below.
In this case, the control means 21 controls the transfer means 17 so as to transfer the accommodating plate 3 supported by the third support means 16 to the first support means 14 and moreover, the transfer means 17 places the different-kind accommodating plate 3b supported by the second support means 15 onto the elevating table 24.
In that state, the engaging claw 15a of the second support means 15 is retracted, the elevating table 24 is lowered, and the accommodating plate 3b is transferred to the third support means 16. In this state, similarly to the case of the cell mass 1 accommodated in the accommodating plate 3, the different-kind cell mass 1b accommodated in the accommodating recess portion 3a of the different-kind accommodating plate 3b is processed.
Moreover, if the cell masses 1 are taken out of all the accommodating recess portions 3a in the accommodating plate 3, the transfer means 17 transfers the empty accommodating plate 3 supported by the third support means 16 to below the recovery magazine 13 and accommodates it in the recovery magazine 13.
Number | Date | Country | Kind |
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2011-140111 | Jun 2011 | JP | national |
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/JP2012/065568 | 6/19/2012 | WO | 00 | 12/16/2013 |
Publishing Document | Publishing Date | Country | Kind |
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WO2012/176751 | 12/27/2012 | WO | A |
Number | Name | Date | Kind |
---|---|---|---|
20050026221 | Richmond | Feb 2005 | A1 |
20080063572 | Deutsch | Mar 2008 | A1 |
20080257074 | Fuchiwaki | Oct 2008 | A1 |
20110200559 | Koga | Aug 2011 | A1 |
Number | Date | Country |
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2006-141326 | Jun 2006 | JP |
2006-271210 | Oct 2006 | JP |
4517125 | May 2010 | JP |
WO 2008123614 | Oct 2008 | WO |
Entry |
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English translation of International Search Report for PCT/JP2012/065568 (1 page). |
Number | Date | Country | |
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20140120192 A1 | May 2014 | US |