JPH0637678Y2 - Cell-shaped cell electrical stimulation chamber - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial field of application) The present invention relates to a cell-shaped chamber used in a cell electrical stimulator for fusing cells with each other or introducing genes into cells by utilizing electrical stimulation. It is a thing.

(Prior Art) There are various types of chambers used in a cell electrical stimulation apparatus. As an example of the chamber, as shown in FIG. 9, there has been proposed one in which a pair of electrodes 2 and 3 are pasted with an adhesive so as to face the inner surface of a rectangular tubular cell 1 made of a resin and having a bottom. There is. Aluminum foil is used as the electrodes 2 and 3, for example. 7 is a cell suspension contained in the cell 1, to which a voltage is applied by the electrodes 2 and 3.

(Problems to be solved by the device) Since the electrodes 2 and 3 are attached to the inner wall surface of the cell 1, there is a gap between the electrodes 2 and 3 and the bottom surface 5 at the bottom as shown in FIG. There are things you can do. In particular, when the cells 8 are precipitating cells, electrical stimulation is given in the state where the cells 8 are deposited on the bottom surface 5, so that if a gap 6 is formed between the tips of the electrodes 2 and 3 and the fusion efficiency is improved. Gene transfer efficiency is reduced. 4 is a line of electric force.

Further, the cell type chamber shown in FIG. 9 is used by being placed vertically as shown in the figure. Depending on the type of cell, the sedimentation speed is fast, and cell fusion and gene transfer are performed in the state of precipitating at the bottom, but the vertical cell-shaped chamber has a small bottom area, so the density of the cells precipitating at the bottom is too high and the fusion occurs. There is also a drawback that gene transfer is not successful.

The present invention makes it possible to perform cell fusion and gene transfer by arranging the cell-shaped chamber in the horizontal direction, and to prevent the cell density from becoming excessively high even in the case of precipitating cells, and in a state in which it is placed in the horizontal direction. It is an object of the present invention to provide a chamber in which there is no gap between the electrode and the bottom surface.

(Means for Solving the Problems) Referring to FIG. 1, FIG. 2 and FIG. 3 showing an embodiment, the chamber of the present invention is an integrally molded bottomed rectangular tube. One pair of opposite side walls of one is electrodes 10a, 10b at least a portion of the bottom side is set to a predetermined interval,
The distance between the openings 11a, 11b is set to be wider than the distance between the electrodes 10a, 10b, and one side wall opposite the one side wall 13b on the opening side of the transparent resin side wall 13b. The transparent resin side wall 13b extends between the pair of opening-side portions 11a and 11b.
A partition member 14 lower than the height of the upper facing side walls 11a, 11b is provided, and the bottom surface 12, electrodes 10a, 10b, side walls 11a, 1 thereof are provided.
The cell suspension 7 is accommodated in the space formed by 1b, 13b and the partition member 14 and electric stimulation is given.

(Operation) As shown in FIG. 4, the chamber 15 is placed in the vertical direction to allow the cell suspension to pass from the opening of the partition member 14 to the Pasteur 17
Etc. Then, as shown in FIGS. 1 and 2, the chamber 15 is placed laterally so that the partition member 14 is on the lower side.
The space formed by b, the bottom surface 12, the side wall 13b and the partition member 14 (in the figure, a part of the side wall 11a.11b on the opening side of the electrodes 10a, 10b is also included) is filled.

It is easy to make the area of the side wall 13b, which is sandwiched between the electrodes 10a and 10b and becomes the bottom in the state of being placed in the horizontal direction, larger than the area of the bottom 12 when placed in the vertical direction, which is usually used in the cell type chamber. It is like that.

When the cells are precipitating, they are deposited on the side wall 13b which is the bottom surface, and are subjected to electrical stimulation from the electrodes 10a and 10b for cell fusion or gene transfer.

When the cell suspension 7 is collected after the electric treatment is completed, the chamber is placed in the vertical direction again as shown in FIG.
Collect from the opening with Pasteur 17, etc.

(Embodiment) FIG. 1 is a sectional view showing a chamber of one embodiment seen from above, FIG. 2 is a sectional view seen from the front of the chamber, and FIG. 3 is a right side view of FIG.

10a and 10b are a pair of electrodes provided in parallel, and are made of, for example, aluminum. 11a and 11b are electrodes 10a and 1
0b is an upper side wall on the opening side, 13a, 13b are side walls, and 12 is a bottom surface.The electrodes 10a, 10b, the side walls 11a, 11b, 13a, 13b and the bottom surface 12 form a vertically long rectangular cylindrical chamber having a bottom. ing. The side walls 11a, 11b, 13a, 13b and the bottom surface 12 are made of a transparent resin, for example, a polycarbonate resin, and are integrally formed by using the electrodes 10a, 10b as inserts.

On the opening side, all four surfaces 11a, 11b, 13a, 13b are made of resin,
The distance between the side walls 11a and 11b is wider than the distance between the electrodes 10a and 10b, so that the cell suspension 7 can be easily injected and collected.

On the opening side, as shown in the figure, electrodes 10a, 10b and side walls are provided with a partition member 14 which is vertically erected on a side wall 13b which is a bottom surface when the electrodes 10a, 10b are placed in a horizontal direction so as to be in a vertical direction. 1
It is formed integrally with 3a, 13b, etc. The partition member 14 includes electrodes 10a, 10b, side walls 11a, 11b, 13b and a bottom surface 12.
At the same time, a recess for accommodating the cell suspension 7 is formed. Since the partition member 14 does not cover the entire opening of the chamber 15 but has an opening, the cell suspension 7 is filled in the concave portion determined by the height of the partition member 14 in a state where the chamber 15 is placed in the lateral direction. Can meet.

When cell fusion or gene transfer is performed, the cell suspension 7 is placed in the chamber 15 and placed in the lateral direction as shown in FIGS. 1 and 2, and a power source is connected to the electrodes 10a and 10b for electrical stimulation. give.

Reference numeral 16 is an objective lens of an inverted microscope, and since the side walls 13a and 13b are made of transparent resin, the state of cell fusion and gene transfer can be observed with a microscope.

The chamber of the present invention can of course also be used vertically. In that case, the partition member 14 is not necessary.

When used as a vertical type, in order to prevent invasion of various bacteria into the cell, cover as shown in Fig. 5.
Cover 18 and perform cell fusion and gene transfer. FIG. 6 is a cross-sectional view seen from the front showing the case of using it as a vertical type,
The drawing is a sectional view as seen from the side surface, and FIG. 8 is a sectional view taken along the line AA in FIG.

(Effects of the Invention) The chamber of the present invention has a pair of opposing walls as electrodes, has a partition member having an opening on the opening side, and the other is made of transparent resin, and is integrally molded into a vertically long rectangular cylinder with a bottom. Since it is formed by the method described above and can be used by laying it sideways, it becomes possible to give an electrical stimulation with a larger bottom area than that of a vertical chamber, which is particularly advantageous in the case of precipitated cells.

And, since the space between the side wall and the bottom face when the electrode is laid is integrally molded, it is easy to form the angle between the electrode and the bottom face at a right angle, and There is no gap between them. Therefore, the efficiency of cell fusion and gene transfer is increased, especially in the case of precipitated cells.

When injecting the cell suspension or collecting the treated cell suspension, it is possible to place this chamber in the vertical direction and perform injection and collection operations from the opening with a Pasteur, etc. Excellent recovery operability.

In addition, since the gap on the side of the opening is wider than the gap between the electrodes, injection and recovery are facilitated.

Further, the chamber in which the electrode and the side wall are integrally molded in this way is low in cost and can be a disposable type.

[Brief description of drawings]

FIG. 1 is a cross-sectional view showing a chamber of one embodiment as seen from above, FIG. 2 is a cross-sectional view as seen from the front of the chamber, FIG. 3 is a right side view of FIG. 2, and FIG. FIG. 5 is a front view showing a state of injecting a cell suspension into a cell, FIG. 5 is a front view showing a mode in which the chamber is used as a vertical type, and FIG. , FIG. 7 is a cross-sectional view showing the same aspect as seen from the side, and FIG. 8 is AA of FIG.
FIG. 9 is a sectional view showing a conventional cell-shaped chamber, and FIG. 10 is a sectional view showing a bottom portion of the same chamber. 7 ... Cell suspension, 10a, 10b ... Electrode, 11a, 11b, 13a, 13b ... Side wall, 12 ... Bottom, 14 ... Partition member.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Office reference number FI technical display location C12N 15/87

Claims (1)

[Scope of utility model registration request] 1. A pair of opposing side walls of a vertically-elongated bottomed rectangular cylinder integrally formed are electrodes 10a, 10b whose at least bottom side portions are set at a predetermined interval, and an opening side portion 11a. , 1
The interval of 1b is set to be wider than the interval of the electrodes 10a, 10b, the other pair of opposite side walls is made of transparent resin, and the opening on one side 13b of the transparent resin side walls is formed in this rectangular tube. On the part side, a portion 11a on the opening side of the pair of opposite side walls,
A partition member 14 extending between 11b and having a height lower than the height of the opposing side walls 11a and 11b on the transparent resin side wall 13b is provided in a state where this rectangular tube is placed so that the transparent resin side wall 13b is located below. , Bottom surface 12, electrodes 10a, 10b, side walls 11a, 11b, 13b
A cell-shaped cell electrical stimulation chamber in which a cell suspension 7 is housed in a space constituted by a partition member 14 and electrical stimulation is given.