JP4249463B2 - Human tissue culture device - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a culture device for human tissue applied in the medical field. In place Related.
[0002]
[Prior art]
Obedience Come For example, when a tear or damage of a tendon or a ligament occurs, these tissues are joined together. For example, autologous transplantation for transplanting other tissues of the living body and the transplanted tissue may be reinforced with artificial materials. Alternatively, an artificial substitute is used as a material for linking tissue torn or damaged parts. Such a connecting material is formed by weaving or knitting synthetic fibers such as polyester and polypropylene that are non-degradable to a living body. In addition, since such a material is non-absorbable to a living body and remains semi-permanently in the living body, if any problem occurs, it may have to be removed by re-operation.
[0003]
[Patent Document 1]
Japanese Patent Application No. 2001-318503
[0004]
[Problems to be solved by the invention]
In general, the connecting material as described above is preferably a material having physical properties such as strength, torsional characteristics, and elongation characteristics that are close to those of a living body. Furthermore, from the viewpoint of tissue regeneration, those having properties closer to that of a living body, for example, those having cells attached to a substrate are desirable. This is because the affinity between the connecting base material and the living body is increased in a state where the connecting material is retained in the living body as described above. However, it is difficult to mass-produce a connecting material formed by attaching cells to a base material so that it can withstand the demands of the medical field, and to manage the quality so as to meet a desired standard.
[0005]
The present invention relates to a culture apparatus for human tissue capable of mass-producing a connecting material formed by attaching cells to a base material with a desired quality. Place An object of the present invention is to provide a substrate closer to a living tissue by providing, that is, by repeatedly applying tension, particularly after cells are adhered.
[0006]
[Means for Solving the Problems]
The apparatus for culturing human tissue according to the present invention comprises driving means for applying tension to the base material immersed in the medium, and a sealed culture unit for storing the culture medium and the base material, wherein the driving means is used for the closed culture. It arrange | positions on the outer side of the part, It is characterized by the above-mentioned.
[0007]
Furthermore, the culture apparatus for human tissue according to the present invention provides a medium tank for storing the medium, and tension is applied to the substrate based on the driving force of the driving means while holding the substrate inside the medium tank. A culture unit including a chuck device is detachably attached to the sealed culture unit.
[0008]
Furthermore, in the human tissue culturing apparatus according to the present invention, the chuck device is capable of advancing and retreating with respect to the stationary chuck, with the stationary chuck sandwiching one end of the substrate and the other end of the substrate. The movable chuck is moved forward and backward by transmitting a driving force to the movable chuck from the driving means via the transmission mechanism.
[0009]
Furthermore, in the human tissue culturing apparatus according to the present invention, the transmission mechanism includes an adjuster mechanism that adjusts a stroke in which the movable chuck moves back and forth.
[0010]
Furthermore, in the human tissue culture apparatus according to the present invention, the fixed chuck and the movable chuck can be tilted in a direction in which the substrate is twisted with respect to each other.
[0011]
Furthermore, the human tissue culture apparatus according to the present invention comprises a load measuring means for measuring the tension applied to the substrate.
[0012]
Furthermore, in the human tissue culture apparatus according to the present invention, the transmission mechanism is configured to transmit a driving force from the driving means to the movable chuck via an elastic body that expands and contracts in the advancing and retreating direction of the movable chuck. It is.
[0013]
Furthermore, the human tissue culture apparatus according to the present invention is configured such that the elastic body is detachably attached to the transmission mechanism.
[0014]
Furthermore, the human tissue culture apparatus according to the present invention includes the adjuster mechanism including a cam that is rotated by the drive source, and a swing link that moves the movable chuck forward and backward by being driven by the cam. By making the amount of eccentricity variable, the stroke of the swing link can be expanded and contracted.
[0015]
Furthermore, the human tissue culture apparatus according to the present invention can control the tension applied to the base material by selecting the stroke of the swing link and the elastic coefficient of the elastic body.
[0016]
Furthermore, in the human tissue culture apparatus according to the present invention, the drive source is a variable speed motor having an output shaft connected to the cam.
[0017]
Furthermore, the human tissue culturing apparatus according to the present invention has a period based on the rotation speed of the output shaft of the variable speed motor, and tension is repeatedly applied to the base material, thereby increasing or decreasing the rotation speed of the output shaft, The period can be adjusted.
[0018]
DETAILED DESCRIPTION OF THE INVENTION
Human tissue culture apparatus according to an embodiment of the present invention In place explain about. A partially broken side view of the human tissue culture apparatus according to the present embodiment is shown in FIG. 1, and the plan view of the apparatus is divided into FIGS. 2 and 3, respectively.
[0019]
As shown in FIGS. 1 to 3, the human tissue culture apparatus 1 according to the present embodiment includes a plurality (two in the drawing) of medium tanks 3 that store the medium 2 in the bottom, and a plurality (four in the drawing). A chuck device 5 that holds the base material 4 in the culture medium 2 while being immersed in the culture medium 2, and a sealed culture unit 6 that houses the culture medium 2, the culture medium tank 3, the base material 4, and the chuck device 5. The drive unit 7 applies tension to the individual base material 4 from the outside of the closed culture unit 6.
[0020]
The medium 2 is a culture solution composed of a cell suspension or a cell culture solution. The culture medium tank 3 is a sealed container made of rectangular acrylic resin or the like that can be divided into two vertically and can be opened and closed by attaching and detaching the upper part to the lower part. As shown in FIG. 2, two bases 4 are stored in two medium tanks 3. As the base material 4, a braid or a thread can be applied. The sealed culture unit 6 is a well-known incubator.
[0021]
In addition, all the base materials 4 may be stored in one medium tank 3, or four base materials 4 may be stored in four medium tanks 3 one by one. The number of the culture medium tank 3, the base material 4, the chuck device 5, or the driving means 7 described above can be increased or decreased as appropriate.
[0022]
The chuck device 5 is disposed inside the medium tank 3 and has a plurality of fixed chucks 50 sandwiching one end of the base material 4. The chuck device 5 is disposed inside the medium tank 3 so as to face these fixed chucks 50. It comprises a plurality of movable chucks 51 sandwiching the other end.
[0023]
The movable chuck 51 moves forward and backward in a direction away from or approaching the fixed chuck 50 by the driving force transmitted from the driving means 7 via the plurality of transmission mechanisms 9. The strokes at which these movable chucks 51 advance and retreat can be individually adjusted by a plurality of adjuster mechanisms 10. Details of the transmission mechanism 9 and the adjuster mechanism 10 will be described later.
[0024]
As shown in FIGS. 1 and 4, the fixed chuck 50 fixes an L-shaped member 50 b with a screw 50 c in the vicinity of the tip of the shaft portion 50 a that passes through a proper position at one end of the medium tank 3. One end of the base material 4 is sandwiched between the horizontal portion 50d and a square seat 50f fixed with a screw 50e in a posture facing the horizontal portion 50d. The upper end of the L-shaped member 50b is divided into two forks, and the shaft portion 50a is sandwiched therebetween. By tightening the screw 50c, the upper end of the L-shaped member 50b can be fixed to the shaft portion 50a. If the screw 50c is loosened, the position of the L-shaped member 50b can be adjusted by appropriately moving in the thrust direction with respect to the shaft portion 50a.
[0025]
The thrust direction of the shaft portion 50a is the same as the direction in which the movable chuck 51 moves back and forth. Hereinafter, this is referred to as an “advance / retreat direction”.
[0026]
The movable chuck 51 is obtained by fixing an L-shaped member 51b facing the L-shaped member 50b in the vicinity of the tip of the shaft portion 51a penetrating the position corresponding to the shaft portion 50a at the other end of the medium tank 3. The L-shaped member 51b and the above-mentioned L-shaped member 50b are the same parts (50e and 50f have the same reference numerals) whose directions are different from each other. If the screw 51c is loosened, the position of the L-shaped member 51b can also be adjusted in the forward / backward direction.
[0027]
The base material 4 may be attached to the fixed chuck 50 and the movable chuck 51 in a twisted state. The angle at which the substrate 4 is twisted can be set in a stepless manner to be, for example, 30 °, 60 °, or 90 ° by exchanging the parts of the movable chuck 51. That is, instead of the L-shaped member 50b which is a component of the movable chuck 51, a rotating L-shaped member 52 as shown in FIG. 5 is applied.
[0028]
The rotating L-shaped member 52 includes a joint member 52b fixed with a screw 52a in the vicinity of the tip of the shaft portion 51a, and a U-shaped sandwiching member 53 attached to the lower end portion of the joint member 52b. The joint member 52b has an insertion hole 52c penetrating the lower end portion in the advancing / retreating direction, and an angle fixing screw 52d is screwed into a screw hole penetrating the lower end portion in a direction perpendicular to the insertion hole 52c. Moreover, the upper end of the joint member 52c is divided into two forks, and can be fixed to the shaft portion 51a by tightening a screw 52a. This is the same as the L-shaped members 50b and 51b.
[0029]
The U-shaped sandwiching member 53 forms a notch 53a into which the base material 4 is inserted between two pieces overlapped with each other, and the base material fixing screw 53b is screwed into a screw hole penetrating from the one piece to the notch 53a. Match. The base material 4 can be clamped between the base material fixing screw 53b and the other piece of the U-shaped clamping member 53 by tightening the base material fixing screw 53b. If the base material fixing screw 53b is loosened, the base material 4 can be freely inserted into and removed from the cut portion 53a.
[0030]
At the end of the U-shaped sandwiching member 53 opposite to the notch 53a, a support shaft 53c is rotatably inserted into the insertion hole 52c of the joint member 52b. A washer 53d is fixed with a screw 53e. By loosening the angle fixing screw 52d and appropriately rotating the U-shaped sandwiching member 53, the U-shaped sandwiching member 53 is aligned so that the base member 4 is twisted at a desired angle. If the angle fixing screw 52d is tightened in this state, the U-shaped clamping member 53 can be fixed to the joint member 52c, and the U-shaped clamping member 53 can be set to a desired angle.
[0031]
In the above description, once the angle of the U-shaped sandwiching member 53 is set, the movable chuck 51 moves forward and backward while maintaining this angle. However, as the movable chuck 51 moves forward and backward, the U-shaped sandwiching member 53. The angle (the angle of rotation about the support shaft 53c as a pivot) may be changed every moment. For example, the U-shaped clamping member 53 is repeatedly rotated clockwise and counterclockwise by a servo motor or the like, and continuously rotated like a pendulum.
[0032]
Further, the fixed chuck 50 and the movable chuck 51 constitute a culture unit 8 together with the medium 2 and the medium tank 3 described above.
[0033]
That is, as shown in FIGS. 1, 4 and 6, the culture unit 8 fixes the vertical frame member 82 to the upper surface of one end of the horizontally long main frame 80, and the linear motion guide bearing 83 a to the upper surface of the other end of the main frame 80. A slider 83 is provided so as to be slidable in the forward and backward directions. The rear end of each shaft portion 50a protrudes outward from one end of the culture medium tank 3, and is supported by the vertical frame member 82 of the main frame 80 via a plurality of bushings 81 through which these are inserted. The rear ends of the individual shaft portions 51 a protrude outward from the other end of the medium tank 3 and are connected to the slider 83 via the clamp 91. On the outer surface of the other end of the medium tank 3, a bellows 30 for sealing the gap between the medium tank 3 and the shaft portion 51a is provided.
[0034]
The clamp 91 rotates the handle 91a to fasten the clamp 91 itself to the shaft portion 51a or allow the clamp 91 itself to slide in the forward / backward direction with respect to the shaft portion 51a. The position adjustment of the entire movable chuck 51 in the advancing / retreating direction, that is, the increase / decrease of the interval between the fixed chuck 50 and the movable chuck 51 is performed by operating the clamp 91 exclusively, that is, relative to the clamp 91 and the entire movable chuck 51. This is done by changing the position.
[0035]
The culture unit 8 is detachably attached to the upper surface of the support base 15 raised inside the closed culture unit 6. Specifically, the hook-shaped claw 16a bent upward is fixed to the main frame 80, and a flange 16b that is hooked on the claw 16a is provided on the horizontal frame 15a that constitutes the upper surface of the support base 15 so as to be swingable. ing. The flange 16b is attached to a short lever 16c that is rotatable about a horizontal axis with a rotation fulcrum at a position slightly below a fulcrum on which it swings.
[0036]
When the short lever 16c is pivoted upward and the short lever 16c is pushed down while the hook 16b is hooked on the claw 16a, the hook 16b can be locked with the hook 16b hooked. If the short lever 16c is pushed upward, the lock can be released. With the above simple operation, the culture unit 8 and the support base 15 can be coupled as shown in FIG. Alternatively, if the culture unit 8 is released from the support base 15, the culture unit 8 can be lifted directly above the support base 15 and removed from the support base 15, as shown in FIG.
[0037]
At the same time as the attachment / detachment of the culture unit 8, the hook-shaped joint 92 fixed to the slider 83 is hooked to the connecting rod 98 of the transmission mechanism 9 so that the culture unit 8 and the transmission mechanism 9 can be immediately connected and disconnected. is doing. Details will be described later.
[0038]
The support base 15 has four legs 15b mainly composed of upright screw rods screwed in the vicinity of the four corners of the horizontal frame 15a which is rectangular when viewed from above. By appropriately rotating the leg 15b, the entire horizontal frame 15a can be moved up and down to adjust the height position for holding the culture unit 8, or the culture unit 8 held by the horizontal frame 15a is horizontal. In addition, the delicate inclination of the horizontal frame 15a can be corrected. At the lower end of the leg 15b, there is provided a bottom tread 15c with a thrust bearing or the like for receiving a load applied thereto.
[0039]
Furthermore, a load measuring means 11 is provided at the rear end of each shaft portion 50a protruding outward from one end of the medium tank 3. The load measuring means 11 measures the tension applied to the base material 4 via the fixed chuck 50. That is, the hanging member 50g is fixed to the rear end of the shaft portion 50a, and the pressure receiving portion 11a of the load measuring means 11 is in contact with the hanging member 50g. When tension is applied to the base material 4, this tension is applied to the pressure receiving portion 11a as pressure in the opposite direction via the shaft portion 50a and the hanging member 50g. Accordingly, the pressure is measured by the strain gauge built in the load measuring means 11. This pressure value is nothing but the magnitude of the tension applied to the substrate 4.
[0040]
As shown in FIGS. 1 and 3, the drive means 7 includes a transmission mechanism 9, an adjuster mechanism 10, a drive source 12, and a support base 17.
[0041]
The support base 17 is one in which one end of the top plate 17a is supported by a leg 17b and the other end is supported by a leg 17c. The legs 17b and 17c fix the top plate 17a above the surface plate 18 so that the top plate 17a is substantially the same height as the horizontal frame 15a. The sealed culture unit 6 and the support base 17 are respectively fixed to the upper surface of the surface plate 18 adjacent to the driving means 7. The surface plate 18 is provided with screw jacks 18a at appropriate positions on the lower surface thereof.
[0042]
The transmission mechanism 9 includes a drive pulley 12b fixed to the output shaft 12a of the drive source 12, a driven pulley 12d connected to the drive pulley 12b via an endless flat belt 12c, a camshaft 13a connected to the driven pulley 12d, A plurality (four in the figure) of cams 13 fixed at predetermined intervals in the thrust direction of the camshaft 13a, and a plurality of swing links 14 that are driven by the respective cams 13 to reciprocate the top pin 14a in the forward and backward directions. And a plurality of buffer portions 93 connected to the swing link 14 via a plurality of universal joints 19.
[0043]
The drive source 12 is a DC motor with a speed reducer fixed to the upper surface of the surface plate 18. By increasing or decreasing the voltage supplied to the drive source 12, the rotational speed of the output shaft 12a of the drive source 12 can be freely adjusted. In addition, an AC motor, a rotary solenoid, or the like may be applied as the drive source 12.
[0044]
The predetermined interval related to the cam 13 means an equal interval corresponding to the interval between the plurality of base materials 4 (FIG. 2) arranged side by side in a direction orthogonal to the advancing / retreating direction. The camshaft 13a extends in a direction orthogonal to the forward / backward direction, and both ends thereof are respectively supported by two legs 17c that form a pair in a direction orthogonal to the forward / backward direction.
[0045]
The configuration of the cam 13 includes the adjuster mechanism 10. That is, the cam 13 has a disc-shaped flange 13b having a contour concentric with the axis of the camshaft 13a fixed to the circumferential surface of the camshaft 13a, and an annular cam body 13c inserted through the camshaft 13a. It is arranged so as to be in sliding contact with. On the other hand, the block piece 10a penetrating the screw hole is fixed to the surface of the cam body 13c on the flange 13b side with a screw 10b, and the adjusting bolt 10c is screwed into the block piece 10a so as to protrude toward the outer peripheral surface of the flange 13b. ing.
[0046]
By appropriately rotating the adjusting bolt 10c, the dimension of the adjusting bolt 10c protruding from the block piece 10a can be increased or decreased. This dimension defines the amount of eccentricity (the lift amount of the cam 13) between the axis of the camshaft 13a and the axis of the cam body 13c. With the eccentric amount adjusted to a desired value, the flange 13b and the cam body 13c are fixed by tightening a screw 13d that passes through the cam body 13c and is screwed into the flange 13b. A handle 13d is provided at the end of the camshaft 13a opposite to the driven pulley 12d so that the camshaft 13a can be manually rotated.
[0047]
The adjuster mechanism 10 and the cams 13 described above are incorporated in the culture apparatus 1 by the number of the base materials 4, that is, four in the drawing. This is because the dimensions and physical properties of the individual base materials 4 are subtly different from each other, and in order to apply equal tension to the individual base materials 4, it is essential to adjust the four adjuster mechanisms 10 separately. It is. Further, the individual adjuster mechanisms 10 can be adjusted while actually measuring the tension actually applied to each base material 4 by a plurality (four in the drawing) of load measuring means 11.
[0048]
The swing link 14 has one end provided with the top pin 14a facing upward, the lower end provided with the driven roller 14b facing downward, and a position that bisects the space between the top pin 14a and the driven roller 14b. The inserted support shaft 14c is turned around a fulcrum. The support shaft 14c extends in parallel with the camshaft 13a, and both ends thereof are respectively supported by two legs 17c. A return spring 14 d is provided near the upper end of the swing link 14. The return spring 14 d urges the swing link 14 with its elastic force and presses the driven roller 14 b toward the cam 13.
[0049]
The driven roller 14b comes into contact with the outer peripheral edge of the cam 13 and serves as a contact point between the swing link 14 and the cam 13. However, the driven roller 14b is disposed on one side of the swing link 14, and accordingly The swing link 14 is offset with respect to the cam 13 in the thrust direction of the camshaft 13a.
[0050]
The buffer portion 93 includes a plurality of sliders 94 provided on the upper surface of the top plate 17a of the support base 17 so as to be slidable in the forward and backward directions via linear motion guide bearings 94a, and the upper surface of the top plate 17a adjacent to each slider 94. A plurality of sliders 95 provided so as to be slidable in the forward and backward directions via linear guide bearings 95a, and an elastic body 90 spanned between pins 94b and 95b provided on the plurality of sliders 94 and 95, respectively. Composed.
[0051]
Examples of the elastic body 90 include a coil spring or rubber. In addition, a cylinder type air damper or the like may be interposed between the pins 94b and 95b. As shown in the figure, the coil spring applied as the elastic body 90 is only hooked on the pins 94b and 95b, so that it can be easily attached to and detached from the sliders 94 and 95.
[0052]
Each top pin 14 a of the plurality of swing links 14 is fitted into the input side bearing 19 a of the universal joint 19. On the other hand, each slider 94 has a pin 20 fixed on the upper surface thereof fitted in the output side bearing 19 b of the universal joint 19. The slider 94 advances and retreats according to the reciprocating motion of the top pin 14 a of the swing link 14. Further, the slider 95 connected to the slider 94 via the elastic body 90 also advances and retreats.
[0053]
The slider 95 has a clamp 97 fixed to the upper surface thereof, and a connecting rod 98 is fastened to the clamp 97. The connecting rod 98 protrudes from the driving means 7 and penetrates from the side wall 60 of the sealed culture part 6 to the inside thereof. A guide sleeve 98 a for guiding the connecting rod 98 is embedded in the side wall 60 of the closed culture unit 6. A bellows 61 is provided on the outer surface of the side wall 60 to seal the gap between the connecting rod 98 and the guide sleeve 98a.
[0054]
The clamp 97 allows the clamp 97 itself to be clamped to the connecting rod 98 by rotating the handle 97a, or allows the connecting rod 98 itself to slide relative to the clamp 97 in the forward / backward direction. That is, even if the slider 95 is slightly moved as the lift amount of the cam 13 is adjusted, the relative position between the clamp 97 and the connecting rod 98 can be changed accordingly. Therefore, the positions of the connecting rod 98 and the movable chuck 51 can be kept constant regardless of the change in the lift amount of the cam 13.
[0055]
As shown in FIG. 7A, a T-shaped flange 98b that protrudes in a direction orthogonal to the advancing / retreating direction is formed at the end (tip) of the connecting rod 98 on the slider 83 side. On the other hand, the hook-shaped joint 92 is fixed to the slider 83 with screws 92a in a state of being paired with a distance through which the connecting rod 98 can be inserted. When the culture unit 8 is lifted straight up when the culture unit 8 is removed from the support base 15 as shown in FIG. 6, the pair of hook joints 92 are connected to the connecting rod as shown in FIG. 7 (b). It easily disengages from the 98 flange 98b.
[0056]
In addition, the connection and disconnection work between the slider 83 and the connecting rod 98 is quick. Or As long as it is a joint that can be easily performed, it is not limited to the above configuration. For example, in a state in which the slider 83 and the connecting rod 98 are overlapped with each other, the two may be separated by penetrating them with a pin orthogonal to the advancing / retreating direction and extracting the pin. .
[0057]
The sliders 94 and 95 are arranged in a line in the forward / backward direction, and the linear motion guide bearings 94a and 95a share a single guide rail. Further, the tension transmitted by the transmission mechanism 9 can be changed by increasing or decreasing the number of the above-described coil springs or the like, or replacing the coil springs or the like with ones having different elastic coefficients.
[0058]
Next, the human tissue culture apparatus according to the present invention Set Examples will be described below.
[0059]
Examples of the artificial human tissue (processed object) manufactured by the human tissue culture apparatus 1 include an exerciser that applies tension to the human body, such as an artificial ligament, an artificial tendon, an artificial rotator cuff, and an artificial muscle. In particular, the human tissue culture apparatus 1 is useful for forming an artificial ligament.
[0060]
As the base material 4, in addition to the in vivo non-absorbable materials described in the prior art, biodegradable and absorbable materials such as lactic acid, glycolic acid, caprolactone, dioxanone, trimethylene carbonate homopolymers, and copolymers thereof Etc., conventional of A polymer used for medical purposes can be appropriately selected and used according to the purpose. In particular, it is desirable to use polylactic acid having high strength and moderate degradability.
[0061]
Examples of the form of the substrate 4 include a knitted fabric, a woven fabric, a non-woven fabric, a thread, or a sponge. In particular, it is desirable to apply a braid excellent in moderate elongation and strength as the base material 4. A braid means a flat braid or a round braid.
[0062]
Examples of the cells cultured in the culture medium 2 in the closed culture unit 6 include fibroblasts, cells constituting ligaments, and mesenchymal cells derived from bone marrow. An appropriate amount of a cell growth factor may be added to these cell culture solutions.
[0063]
A procedure for culturing cells in the medium 2 in the sealed culture unit 6 will be described. That is, one end of the base material 4 having a total length of 30 to 100 mm is sandwiched between the fixed chuck 50 and the other end of the base material 4 is sandwiched between the movable chuck 51, so that the fixed chuck 50 and the movable chuck 51 are sandwiched. The base material 4 is passed over. In this state, the bottom of the medium tank 3 is filled with the medium 2 to such an extent that the base material 4 is immersed in the medium 2. As the medium 2, 100 to 200 ml of a culture solution containing 106 / ml human fibroblasts was used.
[0064]
In addition, after seed | inoculating a cell in the base material 4 beforehand, you may fix the both ends of the base material 4 to the fixed chuck | zipper 50 and the movable chuck | zipper 51 by aseptic operation, respectively. In this way, it is not necessary to put cells in the medium 2 in advance. Moreover, tension can be applied from a state where cells are adhered to the base material 4 in a large amount.
[0065]
The above operation may be performed after the culture unit 8 is detached from the support base 15 and taken out of the sealed culture unit 6. After the culture unit 8 set in this way is returned to the closed culture unit 6 again, the closed culture unit 6 is closed and the temperature inside the closed culture unit 6 is adjusted. Further, when the drive source 12 is activated, the movable chuck 51 moves back and forth and reciprocates, whereby tension is repeatedly applied to the base material 4 spanned between the fixed chuck 50 and the movable chuck 51 at a predetermined cycle. .
[0066]
As specific conditions, it is desirable to set the cycle in the range of about 60 times / minute and the tension in the range of about 4.5 kgf or 10 to 100 N. Such a cycle can be easily set by adjusting the rotational speed of the main shaft 12a of the drive source 12. Such tension can be easily set by adjusting the lift amount of the cam 13 (about 10 mm) by the adjuster mechanism 10 and appropriately selecting the spring constant or elastic coefficient of the elastic body 90.
[0067]
Under the conditions exemplified above, tension is repeatedly applied to the substrate 4 continuously for 2 to 3 weeks. As a result, it was observed that tension was repeatedly applied to the cells attached to the base material 4 and the morphology of the cells changed along the fibers. Moreover, about the base material 4 obtained through the above process, the quantity and quality of the produced | generated collagen are evaluated morphologically and biochemically.
[0068]
It should be noted that the embodiment exemplified above should not be construed as substantially limiting the technical idea of the present invention. In addition, the present invention can be carried out within the scope not departing from the gist of the present invention with appropriate improvements, changes or additions based on the inventive ideas and ideas of those skilled in the art.
[0069]
For example, as the driving source 12, an air cylinder or a solenoid that linearly moves the universal joint 19 or the slider 94 in the advancing / retreating direction may be directly connected. Alternatively, the swing link 14 may be swung around the support shaft 14c by an air cylinder or a solenoid. Further, as a preferable example in workability such as replacement of the culture medium 2, the culture unit 8 is detachable from the support base 15, but the specification may be that the culture unit 8 is fixed in the closed culture unit 6.
[0070]
【The invention's effect】
According to the human tissue culturing apparatus of the present invention, since the driving means is arranged outside the sealed culture part, the sealed culture part and the driving means can be isolated. Thereby, it is possible to reliably prevent the heat generated by the operation of the driving means, mechanical vibration, or dust, bacteria, etc. from adversely affecting the cell culture.
[0071]
Furthermore, according to the human tissue culture apparatus according to the present invention, the culture unit including the culture medium tank and the chuck device is detachably attached to the sealed culture unit. When performing, the culture unit can be easily removed from the sealed culture section. For this reason, since the miscellaneous work such as performing the above-described example operation with the hand inserted in the closed culture unit can be avoided, the above-described example operation can be performed easily and neatly.
[0072]
Furthermore, according to the human tissue culture apparatus according to the present invention, the chuck device includes a plurality of fixed chucks, a movable chuck, a transmission mechanism, and an adjuster mechanism, respectively, so that a plurality of base materials can be set at a time. Therefore, the tension | tensile_strength of the same conditions can be simultaneously provided to a several base material. Or each tension | tensile_strength provided to a several base material can be adjusted for every base material.
[0073]
Furthermore, according to the human tissue culture apparatus according to the present invention, the fixed chuck and the movable chuck can freely rotate in the direction of twisting the base material, so that tension is applied under the condition of reproducing the movement of the human body. be able to.
[0074]
Furthermore, according to the human tissue culturing apparatus of the present invention, the tension applied to the base material by the load measuring means can be measured, so that the processing status can be accurately grasped numerically. Moreover, the tension actually applied to the substrate can be detected. In other words, the base material to which the repetitive tension is applied is gradually extended, and the tension applied to the base material is slightly decreased from the initial set value according to the amount of extension. For this reason, it is always necessary to detect the tension applied to the substrate even during the processing.
[0075]
Furthermore, according to the human tissue culture apparatus according to the present invention, the driving force is transmitted from the driving means to the movable chuck via the elastic body that expands and contracts in the advancing and retreating direction of the movable chuck. Limited by the elastic body. This is because when the drive means forcibly pulls the substrate beyond the maximum length that the substrate can be stretched, this excessive tension can be alleviated by stretching the elastic body, so that the substrate breaks. Can be prevented. In addition, even if the strokes of the plurality of movable chucks are slightly different for each movable chuck, the tension applied to each base material can be made substantially uniform by interposing an elastic body. it can.
[0076]
Furthermore, according to the human tissue culture apparatus according to the present invention, each elastic body is detachably attached to the transmission mechanism. Therefore, if several types of elastic bodies having different elastic body coefficients are prepared, The elastic body can be appropriately replaced so that the tension applied to the material has a desired value.
[0077]
Further, according to the human tissue culture apparatus according to the present invention, the eccentric amount of the cam of the adjuster mechanism is configured to be increased or decreased. It can be adjusted so as not to exceed. Accordingly, appropriate tension can be applied to various substrates having different overall lengths and physical properties.
[0078]
Furthermore, according to the human tissue culture apparatus according to the present invention, the tension applied to the base material can be controlled by selecting the stroke of the swing link and the spring constant or elastic coefficient of the elastic body. The effect of imparting appropriate tension to various base materials having different physical properties can be achieved more remarkably.
[0079]
Furthermore, according to the human tissue culture apparatus according to the present invention, since the drive source is a variable speed motor having an output shaft connected to a cam, the entire operation of the culture apparatus can be electrically controlled and the variable speed can be obtained. By changing the rotation speed of the output shaft of the motor, it is possible to adjust the period for repeatedly applying tension to the substrate.
[0080]
In addition, the speed at which the movable chuck moves back and forth changes depending on the acceleration caused by the cam's unique structure. It can be changed in various ways. Therefore, it is possible to apply a load to the base material such that the heel is pulled by the movement of the living body.
[Brief description of the drawings]
FIG. 1 is a side view, partly broken away, of a human tissue culture apparatus according to an embodiment of the present invention.
FIG. 2 is a plan view of the closed culture unit side applied to the human tissue culture apparatus according to the embodiment of the present invention.
FIG. 3 is a plan view of driving means applied to the human tissue culture apparatus according to the embodiment of the present invention.
FIG. 4 is a side view showing the main part of a closed culture unit applied to the human tissue culture apparatus according to the embodiment of the present invention.
FIG. 5 is a perspective view of a rotating L-shaped member applied to the human tissue culture apparatus according to the embodiment of the present invention.
FIG. 6 is a side view showing a state where the sealed culture unit is removed from the human tissue culture apparatus according to the embodiment of the present invention.
FIG. 7 is a perspective view illustrating a connection state and a disconnection state of a hook-shaped joint and a connecting rod applied to a human tissue culture apparatus according to an embodiment of the present invention.
[Explanation of symbols]
1: Human tissue culture device
2: Medium
3: Medium tank
4: Substrate
5: Chuck device
6: Sealed culture section
7: Driving means
8: Culture unit
9: Transmission mechanism
10: Adjuster mechanism
11: Load measuring means
12: Drive source
13: Cam
14: Swing link
15: Support stand
50: Fixed chuck
51: Movable chuck
90: Elastic body

Claims (8)

A culture medium reservoir for storing a culture medium, a chuck device for holding a plurality of base materials having one end and another end in the culture medium in a state immersed in the culture medium, the culture medium, the culture medium tank, the plurality of base materials, And a culture apparatus for human tissue comprising: a sealed culture unit that houses the chuck device therein; and a driving unit that applies tension to the individual base material from the outside of the sealed culture unit,
The chuck device includes a plurality of fixed chucks that sandwich one end of the base material, and a plurality of movable chucks that sandwich the other end of the base material and can move forward and backward with respect to the fixed chuck. By transmitting a driving force from the driving means to the movable chuck via a plurality of transmission mechanisms, the plurality of movable chucks are moved forward and backward.
The plurality of transmission mechanisms drive the driving means via a plurality of adjuster mechanisms that adjust strokes of the plurality of movable chucks to advance and retreat, and a plurality of elastic bodies that extend and retract in the advancing and retreating directions of the plurality of movable chucks. An apparatus for culturing human tissue , wherein force is transmitted to each of the plurality of movable chucks . The apparatus for culturing human tissue according to claim 1 , wherein the fixed chuck and the movable chuck are tiltable in a direction in which the substrate is twisted with respect to each other. The apparatus for culturing human tissue according to claim 1 or 2 , further comprising load measuring means for measuring the tension applied to the substrate. The apparatus for culturing human tissue according to any one of claims 1 to 3 , wherein the elastic body is detachably attached to the transmission mechanism. The adjuster mechanism includes a cam that is rotated by a drive source, and a swing link that moves the movable chuck forward and backward by being driven by the cam, and is configured so that the eccentric amount of the cam can be increased and decreased. The apparatus for culturing human tissue according to claim 4 , wherein a stroke in which the link advances and retracts can be expanded and contracted. The apparatus for culturing human tissue according to claim 5 , wherein a tension applied to the base material can be controlled by selecting a stroke in which the swing link moves back and forth and an elastic coefficient of the elastic body. The apparatus for culturing human tissue according to claim 5 or 6 , wherein the drive source is a variable speed motor having an output shaft connected to the cam. The human body tissue according to claim 7 , wherein tension is repeatedly applied to the base material at a cycle based on the rotation speed of the output shaft of the speed variable motor, and the cycle can be adjusted by increasing or decreasing the rotation speed of the output shaft. Culture equipment.

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