JP6467735B2 - Reciprocating agitation culture device with medium extraction function - Google Patents

Description

  The present invention can satisfactorily stir the contents of, for example, a culture supernatant containing a product and cells contained in the culture container, and continuously only the medium in the culture container. The present invention relates to a stirring culture apparatus that can be pulled out.

  In a culture vessel, for example, in order to culture a culture such as animal and plant cells and fungus bodies at a high density, while culturing the culture in a culture medium such as a culture solution, a fresh culture medium and waste products are added. It is necessary to replace the medium with the contained medium.

  The extraction of the medium containing the waste in the culture container to the outside of the culture container is conventionally performed, for example, as shown in Patent Document 1, a discharge tube and a pump that pressurizes the culture container. And using the culture apparatus provided with a filtration membrane (filter) that does not pass the culture in the contents of the culture vessel and passes only the culture medium in the discharge tube, and the culture by the pump The inside of the container is pressurized to atmospheric pressure or higher, and the pressure causes the contents in the culture container to be introduced into the discharge pipe, and only the medium in the contents is drawn out of the culture container by the filtration membrane. I am doing so.

Japanese Unexamined Patent Publication No. Hei 8-172956

  However, the conventional culture apparatus has a problem in that a continuous culture medium cannot be extracted because a clogged culture occurs in the filtration membrane.

  The present invention solves these problems and provides a reciprocating stirring culture apparatus having a medium extraction function that can satisfactorily stir the inside of a culture vessel.

In order to achieve the above object, the reciprocating agitation culture apparatus having the medium extraction function of the present application includes a culture container in which a culture and a medium are stored, and the culture container having a stirring blade fixed in the culture container. A drive shaft that extends outward and reciprocates in the axial direction, a pipe hole that is formed in the drive shaft, and from which the medium is drawn out, closes the pipe hole, does not pass the culture, and passes only the medium. A filtration membrane and a medium extraction control means for controlling the extraction of the medium through the pipe hole according to the reciprocating motion of the drive shaft.

  According to the present invention, the contents in the culture vessel can be satisfactorily stirred, and only the medium in the culture vessel can be continuously extracted out of the culture vessel.

It is a longitudinal cross-sectional view of the reciprocating stirring culture apparatus which had the culture medium extraction function of Example 1 of this invention. It is a longitudinal cross-sectional view of the reciprocating stirring culture apparatus which had the culture medium extraction function of Example 2 of this invention. It is a longitudinal cross-sectional view of the reciprocating stirring culture apparatus which had the culture medium extraction function of Example 3 of this invention.

  The example of the form for carrying out the present invention is shown below.

  A first embodiment of the present invention will be described with reference to FIG.

  1 is a reciprocating agitation culture apparatus having a medium extraction function of the present invention, 2 is a culture container of the agitation culture apparatus 1, and the culture container 2 includes, for example, cells and fungi of animals and plants And a culture medium such as a culture solution.

  Reference numeral 3 denotes a drive shaft, which is inserted into the culture vessel 2 through the central opening 4 of the top surface portion 2a of the culture vessel 2.

  The drive shaft 3 is supported by a support portion 5 such as a drive shaft seal portion or a thrust bearing portion provided in the opening 4 so as to be movable up and down. It was connected to a reciprocating drive device (not shown) provided above the surface portion 2a and moved up and down by driving the reciprocating drive device.

  Further, the drive shaft 3 is formed with a pipe hole 3a in the central portion in the axial direction over the entire length thereof, and the pipe hole 3a is closed at the upper end portion of the drive shaft 3 and provided on the upper end side portion. The culture medium was drawn out from the inside of the culture vessel 2 through the pipe hole 3a and the outlet 6 in communication with the outlet 6.

  In addition, the outlet 6 is provided with a medium extraction control means (not shown) for controlling the extraction of the medium. Examples of the medium extraction control means include a pump device for suction, a medium A reverse flow mechanism for making the reverse flow, an on-off valve, and a control device for controlling the pump device, the reverse flow mechanism, the on-off valve and the like by a signal from the reciprocating drive device. Then, the culture medium extraction control means extracts the culture medium in the culture vessel 2 by sucking the outlet 6 in accordance with the reciprocation of the drive shaft 3, or the culture medium is removed. The extraction of various media is controlled such as stopping the extraction or applying a positive pressure to the outlet 6 to reversely flow the medium extracted from the pipe hole 3a b. To.

  In the culture medium extraction means, the open / close valve is omitted, and the culture medium is extracted or the extraction of the culture medium is stopped or the culture medium is reversely flowed only by the operation / stop of the pump device. It is also possible to control the extraction of the medium.

  Alternatively, in the culture medium extraction control means, the backflow mechanism may be omitted, and control may be performed only to stop the extraction of the culture medium or the extraction of the culture medium.

  Alternatively, even when the backflow mechanism is omitted, the on-off valve is opened and the negative pressure associated with the reciprocating motion of the drive shaft 3 is used so that the culture medium flows back naturally. The medium extraction control means may control the extraction of the medium such as extracting the medium, stopping the extraction of the medium, or reversing the medium.

  Reference numeral 7 denotes a filtration membrane (filter), and the filtration membrane 7 is attached to the lower end of the drive shaft 3 so as to close the lower end opening of the pipe hole 3a of the drive shaft 3. The filtration membrane 7 is a porous membrane that can pass only the culture medium without passing through the culture in the contents in the culture vessel 2, for example, a membrane made of a material such as fluororesin or cellulose acetate. There is a filter.

  The filtration membrane 7 may be provided in the pipe hole 3a instead of being provided at the lower end of the pipe hole 3a.

  8 is a stirring blade, and the stirring blade 8 is provided at the lower end of the drive shaft 3 in the culture vessel 2, for example, and has a through hole 8a communicating with the pipe hole 3a of the drive shaft 3 at the center. Is formed.

  The stirring blade 8 is provided in the drive shaft 3 in one or more stages, and the position at which the stirring blade 8 is fixed to the drive shaft 3 may be not the lower end but the middle stage. There is no particular limitation.

  Further, the stirring blade 8 has various shapes including a three-dimensional shape such as a circular shape, an elliptical shape, an oval shape, a rectangular shape, a cross shape, and a conical shape. Further, a part or all of the stirring blades may be bent at an arbitrary position.

  As described above, the stirring blade 8 includes various shapes and structures. In the stirring blade 8 of the present embodiment, the blade surface of the stirring blade 8 is in the axial direction of the stirring shaft 3. Are fixed to the stirring shaft 3 at an angle with respect to the vertical plane so that all or part of the blade surface of the stirring blade 8 faces an arbitrary direction. Further, all or a part of the stirring blade 8 is angled around the axis of the stirring shaft 3 (that is, tilted in the axial direction with respect to a line extending radially outward from the stirring shaft 3). Also included are those fixed to the stirring shaft 3.

  Next, the operation of the culture apparatus of the present invention and the effects thereof will be described.

  In the agitation culture apparatus 1 of the present invention, when the drive shaft 3 is reciprocated by the reciprocation drive apparatus, the agitation blade 8 is reciprocated in the vertical direction, so that the contents in the culture vessel 2 (covers) Stirred product) can be stirred well. When the medium in the culture vessel 2 is not extracted, for example, the pump device is stopped and the on-off valve is closed by the medium extraction control means.

  When the culture medium in the culture vessel 2 is to be extracted, when the drive shaft 3 descends when the drive shaft 3 is reciprocated up and down, the culture medium extraction control means controls, for example, the on-off valve. While opening, the said pump apparatus part is operated and the content in the said culture container 2 is attracted | sucked to the said filtration membrane 7 side through the said outlet 6. Then, only the medium in the contents is allowed to pass through the filtration membrane 7 and introduced into the pipe hole 3a, and the medium is extracted out of the culture vessel 2 through the outlet 6. To.

  When the drive shaft 3 is raised, that is, the lower surface 8c of the stirring blade 8 on the outer surface 7a side facing the contents in the culture vessel 2 of the filtration membrane 7 is formed in the culture vessel 2 and the Negative pressure relative to the upper surface 8d on the opposite side of the lower surface 8c (if the inside of the culture vessel is atmospheric pressure, a lower pressure or a lower pressure if the inside of the culture vessel is a desired pressure. The same applies hereinafter. When the drive shaft 3 is moved in the direction of (2)), the medium extraction control means closes the on-off valve, for example, and stops extracting the medium in the culture vessel 2. Furthermore, the medium extracted from the pipe hole 3a may be slightly reversed by the medium extraction control means, for example, by the reverse flow mechanism.

  As a result, even if the culture adheres to the outer surface 7a of the filtration membrane 7 when the drive shaft 3 is lowered, the outer surface 7a of the filtration membrane 7 is moved to the culture vessel 2 when the drive shaft 3 is raised. Since the negative pressure is also applied to the inner surface 7b of the filtration membrane 7 facing the downstream side in the pipe hole 3a, the culture attached to the outer surface 7a of the filtration membrane 7 is separated and clogged. It will be suppressed.

  Further, if the extraction of the medium is stopped and the medium extracted in the pipe hole 3a is slightly reversed, the culture adhered to the outer surface 7a of the filtration membrane 7 is further separated. Clogging will be suppressed.

  Accordingly, the medium extraction control means controls the extraction of the medium so that the medium in the culture vessel 2 is extracted when the drive shaft 3 is lowered, and when the drive shaft 3 is raised, the filtration membrane 7 Since clogging is suppressed, the medium can be continuously extracted by repeatedly controlling the extraction of the medium and the suppression of clogging by the reciprocating motion of the drive shaft 3.

  In the above-described embodiment, the example in which the drive shaft 3 is vertically suspended and the drive shaft 3 is moved in the vertical direction has been described. However, the drive shaft 3 is disposed in the horizontal direction by moving the drive shaft 3 horizontally. You may make it move to an arbitrary direction and to reciprocate in this arbitrary direction. Then, when the drive shaft 3 moves to one side, the medium is extracted, and when the drive shaft 3 moves to the other side that becomes negative pressure, the medium is stopped from being extracted or the medium is slightly reversed. For example, the extraction of the medium may be controlled by the medium extraction control means.

  In addition, when the drive shaft 3 moves to the other side where the negative pressure is generated, it is not necessary to stop or reverse the extraction of the medium by the medium extraction control means, and the drive shaft reciprocates. When the drive shaft 3 moves to the other side after a desired time has elapsed or after a desired number of reciprocations, the medium is extracted by the medium extraction control means. You may make it stop doing or backflow.

  FIG. 2 shows an agitation culture apparatus 9 of Example 2 of the present invention. The agitation culture apparatus 9 uses the agitation blade 10 having no through-hole instead of using the agitation blade 8 having the through-hole 8a in the agitation culture apparatus 1 of Example 1. Further, instead of providing the filtration membrane 7 in the lower end of the pipe hole 3a of the stirring culture apparatus 1 or in the pipe hole 3a, a through hole 11a communicating with the pipe hole 3a is formed in a recess 10a formed on the upper surface of the stirring blade 10. A filter membrane 11 having a desired thickness is fitted and provided, and the stirring blade 10 having the filter membrane 11 on the upper surface side is fixed to the lower end of the drive shaft 3. The inner peripheral surface (inner surface) 11c of the through hole 11a of the filtration membrane 11 faces the pipe hole 3a, and the pipe hole 3a of the drive shaft 3 is set to the inner peripheral surface of the through hole 11a of the filtration membrane 11. 11c and the stirring blade 10 were used.

  The upper surface of the filtration membrane 11 formed the outer surface 11b of the filtration membrane 11 facing the contents of the culture vessel 2.

  In the agitating culture apparatus 9 of Example 2, when the medium in the culture vessel 2 is extracted, the medium is extracted when the drive shaft 3 is lifted when the drive shaft 3 is reciprocated up and down. For example, the control means opens the on-off valve and operates the pump device to suck the contents in the culture vessel 2 toward the filtration membrane 11 through the outlet 6. Then, only the medium in the contents is allowed to pass through the filtration membrane 11 to be introduced into the pipe hole 3a, and the medium is extracted out of the culture vessel 2 through the outlet port 6. To.

  In addition, when the drive shaft 3 is lowered, that is, the upper surface 10b of the stirring blade 10 on the outer surface 11b side facing the contents in the culture vessel 2 of the filtration membrane 11 is formed in the culture vessel 2 and the When the drive shaft 3 is moved in a negative pressure direction with respect to the lower surface 10c opposite to the upper surface 10b, the culture medium extraction control means closes the on-off valve, for example, Stop extracting the medium. As in the first embodiment, the medium extracted from the pipe hole 3a may be slightly reversed by the medium extraction control means, for example, by the reverse flow mechanism.

  As a result, even if the culture adheres to the outer surface 11b of the filtration membrane 11 when the drive shaft 3 is raised, the outer surface 11b of the filtration membrane 11 is attached to the culture vessel 2 when the drive shaft 3 is lowered. Since the negative pressure is also applied to the inner peripheral surface 11c facing the downstream side in the pipe hole 3a of the inner and the filtration membrane 11, the culture attached to the outer surface 11b of the filtration membrane 11 is separated and clogged. Will be suppressed.

  Similarly to Example 1, if the extraction of the culture medium is stopped and the medium extracted in the pipe hole 3a is slightly reversed, the outer surface 11a of the filtration membrane 11 is further increased. The culture adhering to the cell is separated, and clogging is suppressed.

  Accordingly, when the drive shaft 3 is raised, the culture medium in the culture vessel 2 is drawn out, and when the drive shaft 3 is lowered, clogging of the filtration membrane 11 is suppressed. By repeatedly controlling the extraction of clogging and the suppression of clogging, the medium can be extracted continuously.

  FIG. 3 shows the agitation culture apparatus 12 of Example 3 of the present invention. The agitation culture apparatus 12 is the same as the agitation culture apparatus 1 of Example 1, except that the filtration membrane 7 is provided in the recess 8b formed on the lower surface of the agitation blade 8 in place of the lower end of the pipe hole 3a or the pipe hole 3a. A lid having a through-hole 13a communicating with the through-hole 8a of the stirring blade 8 and having a desired thickness of the filtration membrane 13 fitted therein, and closing the through-hole 13a at the center of the lower surface of the filtration membrane 13 14 is provided. The inner peripheral surface (inner surface) 13c of the through hole 13a of the filtration membrane 13 faces the pipe hole 3a, and the pipe hole 3a of the drive shaft 3 is set to the inner peripheral surface of the through hole 13a of the filtration membrane 13. 13c and the lid 14 were closed.

  The lower surface of the filtration membrane 13 formed an outer surface 13b of the filtration membrane 13 facing the contents of the culture vessel 2.

  In the agitation culture apparatus 11 of the third embodiment, when the medium in the culture vessel 2 is extracted, the medium extraction control is performed when the drive shaft 3 is lowered during the reciprocating motion of the drive shaft 3. By means of the means, for example, the on-off valve is opened and the pump device is operated to suck the contents in the culture vessel 2 to the filtration membrane 13 side through the outlet 6. Then, only the medium in the contents is allowed to pass through the filtration membrane 13 to be introduced into the pipe hole 3a, and the medium is extracted out of the culture vessel 2 through the outlet port 6. To.

  In addition, when the drive shaft 3 is raised, that is, the lower surface 8c of the stirring blade 8 on the outer surface 13b side facing the contents in the culture vessel 2 of the filtration membrane is in the culture vessel 2 and the lower surface 8c. When the drive shaft 3 is moved in a negative pressure direction with respect to the upper surface 8d on the opposite side, for example, the open / close valve is closed by the culture medium extraction control means, and the culture medium in the culture vessel 2 is removed. Stop extracting. As in the first embodiment, the medium extracted from the pipe hole 3a may be slightly reversed by the medium extraction control means, for example, by the reverse flow mechanism.

  Thereby, even when the culture adheres to the outer surface 13b of the filtration membrane 13 when the drive shaft 3 is lowered, the outer surface 13b of the filtration membrane 13 is moved to the culture vessel 2 when the drive shaft 3 is raised. Since the negative pressure is also applied to the inner peripheral surface 13c facing the downstream side of the pipe hole 3a in the inner and the filtration membrane 13, the culture attached to the outer peripheral surface 13b of the filtration membrane 13 is separated and clogged. It will be suppressed.

  Similarly to Example 1, if the extraction of the culture medium is stopped and the medium extracted in the pipe hole 3a is slightly reversed, the outer surface 13a of the filtration membrane 13 is further increased. The culture adhering to the cell is separated, and clogging is suppressed.

  Accordingly, when the drive shaft 3 is lowered, the culture medium in the culture vessel 2 is drawn out, and when the drive shaft 3 is raised, clogging of the filtration membrane 13 is suppressed. By repeatedly controlling the extraction of clogging and the suppression of clogging, the medium can be extracted continuously.

  The reciprocating agitation culture apparatus having the medium extraction function of the present invention is used in culture apparatuses for medical products and foods.

DESCRIPTION OF SYMBOLS 1 Stirring culture apparatus 2 Culture container 2a Top surface part 3 Drive shaft 3a Pipe hole 4 Opening part 5 Support part 6 Extraction port 7 Filtration membrane 7a Outer surface 7b Inner surface 8 Stirring blade 8a Through-hole 8b Lower surface 8d Upper surface 9d Stirring culture apparatus DESCRIPTION OF SYMBOLS 10 Stirring blade 10a Recessed part 10b Upper surface 10c Lower surface 11 Filtration membrane 11a Through-hole 11b Outer surface 11c Inner peripheral surface 12 Stirring culture apparatus 13 Filtration membrane 13a Through-hole 13b Outer surface 13c Inner peripheral surface 14 Lid

Claims (5)

A culture vessel in which the culture and medium are contained;
A drive shaft that has a stirring blade fixed in the culture vessel, extends outside the culture vessel, and reciprocates in the axial direction ;
A pipe hole formed in the drive shaft for extracting the medium;
A filtration membrane that blocks the pipe hole, does not pass the culture, and passes only the medium;
A reciprocating agitation culture apparatus having a culture medium extraction mechanism comprising a culture medium extraction control means for controlling extraction of a culture medium through the pipe hole according to the reciprocation of the drive shaft. An opening inside the culture vessel of the pipe hole is provided on the end surface of the drive shaft,
The reciprocating agitation culture apparatus having a medium extraction function according to claim 1, wherein the filtration membrane is provided in the pipe hole or on an end face of the pipe hole . The filtration membrane, the is provided on one surface or the other surface of the stirring blade, said reciprocating agitation culture device having a media extraction function according to the filtration membrane to claim 1 in which to face the said pipe bore . The medium extraction control means, when extracting the medium in the culture container,
When the drive shaft moves in a direction in which the blade surface of the stirring blade on the outer surface side of the filtration membrane facing the contents in the culture vessel becomes negative pressure, the medium in the culture vessel is extracted. A reciprocating agitation culture apparatus having a culture medium extraction mechanism according to claim 1, 2 or 3, wherein the reciprocating stirring culture apparatus is a means for stopping the above. The culture medium extraction control means is means for further backflowing the culture medium in the pipe hole when the drive shaft is moved in a direction in which the blade surface of the stirring blade becomes negative pressure. 5. A reciprocating stirring culture apparatus having the medium extraction mechanism according to 4 .

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