JPH02135087A - Culture device of tissue cell - Google Patents

Abstract

PURPOSE:To obtain a device convenient for culturing tissue cell of external fertilization, etc., by equipping an opening and closing door comprising a transparent plate at the front of a box-shaped operation chamber, a constant- temperature stage at the bottom, clean air supplying means for the stage and a gas feeding means for pH adjustment. CONSTITUTION:An opening and closing door 2 comprising a transparent plate is set at least at the front of a box-shaped operation chamber 1 and a constant- temperature stage 5 on which a culture container 17 is placed is set at the bottom of the operation chamber 1. Air supplying means 14-16 to send clean air from the upper part of the operation chamber 1 are installed. Further the culture container 17 on the constant-temperature stage 5 is equipped with gas feeding means 21-27 to supply a gas for controlling change of pH.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an apparatus for culturing tissue cells, such as in vitro fertilization cells.

[Conventional technology]

Generally, when culturing animal cells, the ambient temperature is maintained at the same temperature as the body temperature of the animal, and the humidity is maintained at a predetermined level to prevent changes in the pH of the culture medium in this constant temperature and humidity atmosphere. For example, CO□ gas (usually CO
3) is supplied.

An example of a conventional culture apparatus of this type is shown in FIG.

That is, a constant temperature layer 52 made of a water jacket is formed on the inner surface of the main body 51 formed of a heat insulating layer, and the culture chamber 53 inside the main body 51 is kept at a constant temperature. In addition, a water reservoir 54 is installed at the bottom of the culture chamber 53, and the water is supplied to the water reservoir 54 from a CO2 gas cylinder 55 and an air pump 56 via a pipe line 57.
O2 mixed gas is supplied to fill the culture chamber 53 with the humidified mixed gas. As described above, the inside of the culture chamber 53 is maintained at an atmosphere of, for example, a temperature of 37° C., a humidity of 90% RH, and a Co□5%. In FIG. 3, reference numeral 58 denotes a temperature sensor, and the operation of the heater 59 is controlled by its detection signal. 60 is a Co2 gas sensor, and the CO2 gas flow rate is controlled by its detection signal. 61.
62 is a flow meter, and 63 is a culture shelf.

[Problem to be solved by the invention]

However, in the conventional device as described above, the main body 51 including the door (not shown) is opaque.
It is not possible to observe the state of the cells inside 3 from the outside, and if you want to observe the progress of the culture, you must open the door and take out the culture container such as a flask. For this reason, opening and closing the door not only destroys the specified atmosphere within the culture chamber, but also exposes the cells taken outside to severe changes in the atmosphere, or contaminates them with bacteria, weakening cell activity and even killing them. Sometimes.

Particularly in the work related to in vitro fertilization, it is necessary to culture sperm obtained from the male body and observe the ever-changing conditions such as the maturation process of the sperm, the maturation process of the egg obtained from the mother's body, and the maturation process of the fertilized egg. In addition, it is not desirable to subject the cells to frequent changes in the atmosphere as mentioned above, since it is also necessary to carry out operations such as fertilization.

This invention was made in order to solve the conventional problems as described above, and it is possible to observe the progress of culture from the outside, and to perform operations on cultured cells without changing the predetermined atmosphere. The purpose is to provide a culture device that can

[Means to solve the problem]

To achieve the above object, the present invention includes a work chamber formed in a box-shaped body and having a predetermined width and depth, an opening/closing door made of a transparent plate provided at least in front of the work chamber, and a work chamber. A thermostatic stage provided at the bottom of the thermostatic stage on which a culture vessel is placed, a gas supply means for supplying gas to suppress changes in pH to the culture vessel on the thermostatic stage, and a gas supply means for supplying clean air to the working room from above. The invention is characterized in that it comprises an air supply means.

〔Example〕

An embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 shows the overall appearance of the culture device according to the present invention.
is a box-shaped main body, and a work chamber S which also serves as a culture chamber and has a predetermined width and depth is formed at an appropriate height position (at a height that allows an operator to work while sitting on a chair). There is. The width and depth are determined by taking into account the number of instruments such as culture containers accommodated in the work room S, the space required for various operations, and the like.

An opening/closing door 2 that moves in the vertical direction is installed at the front of the work room S, and this opening/closing door 2 is made of a transparent plate such as transparent glass. Transparent windows 3 and 4 may be provided on the sides as necessary.

A constant temperature stage 5 having an area occupying almost the entire bottom of the work chamber S is provided. As shown in FIG. 2, this constant-temperature stage 5 consists of a hollow box into which constant-temperature water in a constant-temperature tank 6 flows through a pipe 8 having a pump 7, and the constant-temperature water flowing into the stage 5 The water is returned to the tank 6 via a pipe 9. The temperature of the constant temperature water in the constant temperature stage 5 and constant temperature tank 6 is determined by a temperature sensor 10.
．． 11, and the operations of the heater 12 and the cooling coil 13 are controlled based on the detection signals.

Reference numeral 14 denotes a blower, which sucks air through a preliminary filter 15 and has a sterilizing and sterilizing function.
Clean air is supplied to the work chamber S from above through an EPA filter 16.

Reference numeral 18 denotes a thermostatic covering that covers the culture container 17 placed on the thermostatic stage 5, and the covering 18 is constituted by a water jacket consisting of a double cylinder whose upper end is closed and whose lower end is open.

The water jacket or covering body 18 is made of a transparent material such as transparent glass, into which the constant temperature water in the tank 6 flows through a flexible conduit 19 branched from the conduit 8. It returns to the tank 6 via a conduit 20.

A Co2 gas supply port 27 is provided at the top of the covering 18, and this supply port 27 is connected to the CO2 gas cylinder 21 via a conduit 22a and a flexible conduit 22b. The conduits 22a and 22b include a flow meter 23, a humidifying bottle 24 housed in a heat insulating cylinder 25 on the constant temperature stage 5, and an H
An EPA filter 26 is provided, and humidified and sterilized CO2 gas is supplied to the inside of the cover 18.

Reference numeral 28 denotes a constant temperature stage for a microscope, and this stage 28 is made of a hollow plate having a transparent part 29 in the center. The constant-temperature water flows through a line 30 and returns to the tank 6 via a flexible line 31.

32 is a constant temperature block placed on the constant temperature stage 5, and is designed to maintain the culture solution in the test tube 33 held by this block 32 at a constant temperature.

The mode of use of the above culture device will be explained.

Clean air that has passed through a filter 16 is constantly supplied into the working chamber S, and this air is discharged from the gap between the lower end of the opening/closing door 2 and the bottom of the working chamber S.

A culture container 17 such as a flask with a flat bottom containing tissue cells and a culture solution is placed on a constant temperature stage 5, and a water jacket, that is, a constant temperature covering 18 is placed so as to cover this culture container 17. Ru. In this way, the culture container 17 is maintained at a constant temperature not only by the stage 5 but also by the cover 18, so that its accuracy is good.

On the other hand, humidified and sterilized CO2 gas flows from the Co2 gas cylinder 21 through the pipes 22a and 22b to the covering body 18.
This CO2 gas increases the P of the culture solution.
H change is suppressed.

Tissue cells are cultured under the above atmosphere,
The situation can be visually observed through the transparent opening/closing door 2 and the cover 18.

Further, the following operations are performed on the constant temperature stage 5.

That is, the opening/closing door 2 is opened to the extent that the worker's arm can fit into the work chamber S, and operations such as fertilization of sperm and eggs, replacement of culture fluid, etc. are performed within the work chamber S. At this time, the culture container 17 is kept at a constant temperature by the constant temperature stage 5, and the working chamber S is kept in a sterile state, so that there is no change in the atmosphere of the cells, and there is no room for contamination by bacteria.

Furthermore, when performing microscopic observation of cells, the sample mounting stage is moved into the work chamber S, and the thermostatic stage 28 is placed on the mounting stage, thereby making it possible to observe the cells while maintaining their temperature.

The above embodiments are merely illustrative, and the present invention can be modified in various ways.

〔Effect of the invention〕

As described above, according to the present invention, the front opening/closing door of the working room, that is, the room that also serves as the culture room, is made of a transparent plate, so that the progress of the culture can be observed from the outside, without causing any changes in the atmosphere to the cells. , good culture can be performed.

Furthermore, when performing various operations on cultured cells, it is necessary to partially open the opening/closing door and perform the operations on a constant temperature stage in the work room. Since it is kept clean by the clean air supply means, it is possible to create a good working environment without contamination with germs, etc., and without adversely affecting the cells.

[Brief explanation of the drawing]

FIG. 1 is an overall perspective view showing an embodiment of the present invention, FIG. 2 is a front view showing the inside of the same, and FIG. 3 is a sectional view showing a conventional example.

Claims (1)

[Claims] 1. A work chamber with a predetermined width and depth formed in a box-shaped body, an opening/closing door made of a transparent plate provided at least in front of this work chamber, and a culture container provided at the bottom of the work chamber. It comprises a constant temperature stage to be placed, a gas supply means for supplying a gas that suppresses changes in PH to the culture container on the constant temperature stage, and an air supply means for supplying clean air into the working chamber from above. A tissue cell culture device characterized by: