JPS5950310B2 - Culture container transport device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a culture container transport device used in an apparatus for automatically culturing living tissues and cells.

In all fields such as medicine, biology, pharmacy, and agriculture,
Biological tissue and cell culture techniques are essential basic experimental techniques for conducting research at the cellular level.

However, subculturing biological tissues and cells is technically difficult, and stable culture strains have not been obtained.

However, recently, with the spread of gas culture technology in incubators, that is, culture technology in a specific gas atmosphere, special cells that were previously considered difficult to grow, such as those of the liver, nervous system, and pituitary gland, have become more popular. Even subculture is becoming possible.

The main points of such currently used cell subculture methods are described below.

First, in order to carry out subculture, a predetermined number of cells are diluted with culture medium into a culture container such as a Petri dish, and then injected into a suspended state, and then cultured statically in an incubator maintained in a predetermined atmosphere. do.

In order to check the culture state after a predetermined period of time has elapsed, a predetermined culture container is removed from the incubator and the degree of cell proliferation is examined using a microscope.

When it is confirmed that the target cells have grown to the full capacity of the specified container, the cells are transferred to sterile clean pliers, and the culture solution in the culture container is aspirated with a pipette and discarded. The cells remaining in the container are washed by injecting a buffer solution, and the buffer used for this washing is again aspirated and discarded.

Next, enzymes such as trypsin are injected to release the cells that have implanted and grown on the bottom of the culture container from the culture container.

The cells released from the bottom of the container by the enzyme are separated and collected using a centrifuge.

That is, the enzyme and free cells in a predetermined culture container are transferred to a centrifuge tube, centrifuged, and the supernatant, which is the enzyme, is discarded by suction.

Depending on the cell type, a centrifuge may not be used to separate the enzyme and cells.

In this case, the enzyme is aspirated and discarded immediately before the release of the implanted cells by the enzyme, and the process moves on to the next step.

Next, a culture solution is injected to dispense the cells at a predetermined dilution concentration, the cells are resuspended with a pipette, and the resuspended cell suspension is dispensed in fixed amounts into new culture containers.

After the dilution and dispensing operation has been completed in this way, the culture container is taken out using clean pliers, placed in a container kept at a predetermined atmosphere, and then cultured again.9 However, in the method described above, It has the following disadvantages.

One of them is that in order to check the proliferation state of tissues or cells using a microscope, it is necessary to frequently take out the culture container from the incubator to the outside air.

For this reason, culture conditions may suddenly change when the cultured material is removed from a predetermined environmental condition such as gas atmosphere, temperature, humidity, etc., resulting in subtle changes in the cultured tissue or cells.

Also, because they come in contact with the outside air, they are subject to contamination by bacteria.

In this way, they are directly affected by the effects of changes in environmental conditions and the contamination of various bacteria.

Next, based on the results of the microscopic observation, the technician will perform the above-mentioned cell separation, collection, dilution, and dispensing subculturing manually in clean pliers. direct effects on the affected tissues or cells occur.

In other words, this shows that it is not possible to perform standardized culture under certain conditions with such conventional culture operations.

The tissue or cells cultured will also depend on the experience and skill of the technician.

This means that it is difficult to standardize and unify the culture technology itself.

For this reason, even when conducting research on the same topic, researchers often come to completely opposite conclusions.

Furthermore, in order to train technicians equipped with these cultivation techniques,
Because it requires at least two years, there is an absolute shortage of personnel, and the reality is that researchers are unable to devote their full efforts to their original research, and instead have to devote considerable energy to branch and leaf cultivation techniques. .

From the above 1, we can prevent contamination of outside air by conventional methods, eliminate the influence of human manipulation,
Furthermore, automatic culture apparatuses have been proposed that aim to automatically culture standardized tissues or cells by standardizing and unifying each operation in culture.

In this automatic culturing apparatus, as described above, cells for subculture are diluted with a culture solution and dispensed into each culture vessel in a predetermined number of cells, and statically cultured.

Furthermore, during static culture, it is necessary to occasionally observe the state of cell proliferation and other conditions using a microscope.

Furthermore, when a predetermined number of cells have proliferated, it is necessary to inject an enzyme solution into the cells to release the cells adhering to the bottom of the culture container, perform centrifugation, and then dispense the cells into a new culture container.

All of these operations need to be performed automatically in a room maintained at a constant atmosphere.For this reason, this automatic culture device has a dispensing mechanism, observation microscope, and other machines placed in the aforementioned room. It is designed to perform various operations and observations.

Therefore, a transport device is also required to move culture containers containing cells between machines installed in a room with a constant atmosphere.

The present invention has been made in view of the above points, and it is an object of the present invention to provide a transport device that can reliably transport a culture container with a relatively simple mechanism.

The details of the conveying device of the present invention will be explained below based on the drawings.

FIG. 1 shows an outline of a culture apparatus maintained in a constant atmosphere, including the transport apparatus of the present invention.

In this figure, 1 is a culture chamber whose interior is maintained at a constant atmosphere, 2 is a separation and dispensing operation stage where various operation mechanisms for separating and dispensing cells to be cultured are arranged, and 3 is a cell A culture state detection mounting table 4 is a stationary culture mounting table on which a microscope and other observation equipment are installed to observe the proliferation state and other conditions.

This stationary culture mounting table 4 has a central portion 4a as shown in FIG.
is removed in a circular shape, and in the area where the culture container 5 is placed, a large number of grooves 4b having a width smaller than the diameter of the container are formed radially around the center.

Further, the other mounting tables also have substantially the same shape.

Reference numeral 10 denotes a conveying device of the present invention having a structure as will be described in detail with reference to FIG.

In FIG. 3, reference numeral 11 is a stand fixed to the main body that moves up and down, 12 is a gear whose shaft portion 12a is fitted and held in the center of the stand 11 so as to be rotatable, and 13 is a gear of the gear 12. It is a holding member fitted into the shaft portion 12a, and the arm guide member 14 is fixed and held at the lower end thereof.

Also, on the outer peripheral surface of this holding member 13, a slot) 1 is provided parallel to the axis.
3a is formed and the tip of the pin 15 fixed to the shaft portion 12a of the gear 12 is inserted into this slot 13a, and a female thread 13b is formed inside and threaded into the threaded portion 16 of the vertical movement shaft 16.
It is screwed onto a.

A pinion shaft 17 is rotatably fitted onto the vertical movement shaft 16, and a pinion 18 is fixed to the tip of the pinion shaft.

19 is a rack that is slidably supported within the arm guide member 14 and meshes with the pinion 18; 20 is an arm fixed to the tip of the rack 19; and 21 is a gear that meshes with the gear 12 and is rotated by the arm rotation motor 22. Ru.

A gear 23 meshes with a gear part 16b formed integrally with or fixed to the upper part of the vertical movement shaft 16, and is rotated by the arm vertical movement motor 24.

A gear 25 meshes with a gear 24 fixed to the pinion shaft 17 and is rotated by a motor 26 for moving the arm back and forth.

Reference numeral 27 denotes a storage table having a disk shape and having a large number of radially provided grooves 27a as shown in FIG. 2, 28 a holding shaft holding the storage table 27, and 29 a gear of the holding shaft 28. The storage table is rotated by a motor 30 for rotating the storage table through a gear that meshes with the portion 28a.

Next, the operation of the conveying device of the present invention will be explained.

For example, to explain the case where the proliferation state of cells in each culture container placed on a static culture mounting table is observed, the whole transport device is moved up and down (not shown), and the table 11 is moved along with the table 11 as shown in FIG. The entire device shown in Figure 1 moves up and down.

Then, when the surface of the storage table 27 reaches a height that matches the surface of the stationary culture mounting table 4, the vertical movement of the apparatus 10 is stopped.

Here, if the groove 27a formed in the storage table 27 does not match the groove 4b formed in the stationary culture mounting table 4, the storage table rotation motor 30 is operated to rotate the gear 29 and the holding shaft 28. The storage table 27 is rotated through the storage table 27 so that the two are aligned.

If the arm longitudinal movement motor 26 is driven here, the gear 25 is rotated, and the pinion 18 is rotated via the gear 24 and the pinion shaft 17, thereby moving the rack 19 back and forth, that is, in the horizontal direction in the drawing.

As the rack 19 moves back and forth, the arm 20 also moves back and forth.

By moving the arm 20 back and forth (in this case, moving forward from the position shown in the figure), the arm 20 is positioned at the lower part of the groove 4b of the stationary culture platform 4.

Next, by driving the arm vertical movement motor 24, the gear 23 is rotated, and by rotating the vertical movement shaft 16, the holding member 13 is vertically moved (in this case, raised). 14, rack 19
, the arm 20 fixed to its tip rises together.

As a result, the arm 20 rises through the groove 4b of the mounting table 4 and rises above the surface of the mounting table 4.

Therefore, the culture container 5 placed on the mounting table 4 is placed on the arm 2.
0 and held at a position away from the surface of the mounting table 4.

Furthermore, when the arm 20 is moved backward by driving the arm back and forth movement motor 26 again, the arm 20 moves into the groove 27a of the storage table 27 together with the culture container 5.

If the arm 20 is lowered by driving the arm vertical movement motor 26 again and stopped at the position shown in the figure, the culture container 5 will be moved onto the storage table 27.

Next, by driving the arm rotation motor 22, the arm 20 is moved to the groove where the culture container that needs to be observed is located among the other culture containers placed on the stationary culture mounting table 4. Then, by repeating the same operations as those described above, the culture container is moved onto the storage stand.

By repeating these operations, after all of the culture vessels containing cells to be observed are transferred from the stationary culture mounting table 4 to the storage table 27, the entire transfer device is moved up and down, so that the surface of the storage table 270 is moved from one place to another. It is stopped at a height that coincides with the surface of the mounting table, in this case the culture state detection mounting table 3.

When the transport device has come to the position of the culture state detection mounting table 3 in this way, by performing a combination of the above-mentioned rotation, vertical movement, and back and forth movement of the arm 20, the culture container containing the cells to be observed is removed. The cells are sequentially moved to the culture state detection mounting table 3 and observed using an appropriately configured observation device (not shown), and after the observation, the cells are returned to the storage table 27 again.

In this way, after all the cells to be observed have been observed, the transport device is moved again to the position of the stationary culture platform 4, and the culture container on the storage platform 27 is placed on the stationary culture platform 4. The cells are then transferred and statically cultured.

The above explanation has been made regarding the case of observing cells in static culture, but basically the same operation can be used when dispensing cells grown by static culture, for example.

In that case, it goes without saying that the conveyance device is moved to the separation/dispensing operating table 2, and the dispensing etc. are performed by an appropriate dispensing device installed on the table 2.

As explained above, according to the transport device of the present invention, all the movement of the culture vessels is carried out within the culture chamber where a constant atmosphere is maintained at all times, so there is no risk that the cells will be contaminated with germs etc. during transport.

In addition, the transport device can be operated by moving the entire device up and down, rotating the storage stand, and by simply moving the arm, so the device is simple and has the advantage of being easy and reliable to operate. has.

[Brief explanation of the drawing]

FIG. 1 is a diagram schematically showing a culture chamber equipped with the transport device of the present invention, FIG. 2 is a plan view showing a part of the mounting table in the culture chamber, and FIG.
The figure is a sectional view of the device of the present invention. 1...Cultivation room, 2...Separation/dispensing operation mounting table, 3...
・Culture state detection mounting table, 4...Static culture mounting table, 5.
...Culture container, 10...Transfer device, 2o...Arm, 27...Storage stand.

Claims (1)

[Claims] 1. A disk-shaped storage table having a large number of radially formed grooves and arranged so as to be rotatable, and a longitudinal direction and a circumferential direction of the grooves of the storage table in the vertical direction and on a horizontal plane. 1. An apparatus for transporting a culture container, comprising an arm configured to be able to move to a position, and a culture container on a storage stand being moved by each movement operation of the arm.