JPH01165366A - Cell fusion chamber - Google Patents

Abstract

PURPOSE:To make it possible to carry out cell fusion without using alternating voltage and centrifugal force, by providing a lower electrode provided at the bottom of a chamber main unit and upper electrode having holes into which a cell can be passed and capable of moving to the state capable of coming into contact with the lower electrode. CONSTITUTION:A lower electrode 6 is provided at the bottom of a chamber main unit and a nylon mesh having insulation properties is bonded thereon. An upper electrode 5 bonded by mesh 7 consisting of similar material is arranged opposite the lower electrode 6. Namely, when a call suspension is charged into the chamber main unit 1, a liquid is floated on the lower electrode 6. Then when the upper electrode 5 is sunk, a cell is pressed between electrodes by self-weight of the upper electrode and simultaneously the liquid is passed into the hole of upper electrode and escaped upward. Then cell fusion is carried out by applying direct current pulse when the cell is pressed. As a result, the cell fusion can be carried out using only direct current pulse generator without using centrifugal separator as well as alternating current source.

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application The present invention relates to a chamber of a cell fusion device that fuses cells using electrical stimulation.

(b) Conventional technology In conventional cell fusion chambers, a cell suspension containing cells is placed between a pair of electrodes, and an alternating current voltage is applied to the opposite electrode to electrophores the cells. Let them touch and then a pair of 1! By applying high-voltage direct current pulses from the poles, cells are electrically stimulated and fused.

In addition, as shown in Fig. 2, another type of cell fusion chamber is a cell fusion chamber in which cells are brought into close contact with each other by centrifugal force acting in the direction of arrow 56 and then fused by applying a DC pulse. Proposed.

In the cell fusion chamber shown in FIG. 2, a lower chamber [52 is provided on the inner bottom surface of the chamber body 50 housed in a centrifuge container, and a lower chamber [52] is provided at a short distance from the chamber body 50 [WS2].
An upper electrode 54 is provided along the inner wall of.

The inner diameter of the chamber body 50 is about 2ff.

When a cell suspension is placed in the chamber body 50 and a centrifugal force is applied in the direction of the arrow 56, the cells are collected on the lower electrode 52, so a voltage is applied between the lower electrode 52 and the upper electrode 54 to cause cell fusion. can be caused to occur.

(c) Problems to be solved by the invention In the former chamber, which applies an alternating current voltage and causes cells to electrophores and come into contact with each other, 1) cells are suspended in a solvent with a high salt concentration, such as seawater; If the electrical resistance of the cell suspension is too low to apply a predetermined voltage to the cell suspension, therefore, after diluting the lfl cell suspension to increase the electrical resistance value, is applied to perform cell fusion.

Some cells may die if the components of the cell suspension are changed. For example, the cells of seaweed and seaweed die when the concentration of seawater changes. Therefore, such cells have been difficult to fuse with conventional devices.

Further, in the cell fusion chamber shown in FIG. 2, the electrodes are small and the lines of electric force 58 are concentrated in the center of the lower electrode 52, making it impossible to obtain parallel lines of electric force, so there is a problem that the amount of cells to be processed is small. be.

An object of the present invention is to provide a cell fusion chamber that allows cells to come into contact without using alternating current voltage or centrifugal force.

B) Means for Solving the Problems The present invention provides a lower electrode provided at the bottom of the chamber body;
It is characterized by comprising an upper electrode which has a hole through which cells can pass and which can be moved to a state where it can be brought into pressure contact with the lower electrode.

Here, the lower N, pole and upper electrode are made of a conductive metal material having an insulating mesh and fine holes.

Examples of conductive metal materials with fine pores include sintered metal (pore diameter 5 μm to 10 μm), and examples of insulating mesh include nylon mesh (pore diameter 10 μm).
0 μm to 200 μm).

(e) When a cell suspension is placed in the action chamber body, the liquid floats on the lower electrode. Next, when the upper electrode pupil is lowered, the weight of the upper electrode presses the multiple cells between the xi and the liquid escapes upward through the hole in the upper electrode. When the cells are pressed together, a direct current pulse is applied to perform cell fusion.

(f) Example An example of the present invention is shown in FIG. The chamber body consists of a resin body 1, and a terminal 4 is screwed into the bottom of the body 1. The lower electrode 6 is placed on top of this.
is made of sintered metal. An insulating nylon mesh 8 is adhered onto the lower electrode 6.

Further, an upper electrode 5 to which a mesh 7 made of a similar material is adhered is arranged facing the lower electrode 6.

Reference numeral 11 denotes a tube, which electrically connects the terminal 3 adhesively attached to the lid 2 and the upper electric pupil 5.

In the cell fusion chamber configured as above.

The fusion is performed as follows.

First, from the bottom of body 1, lower mesh 8. After building up the lower electrode 6 and lower terminal 4, the cell suspension 10 is injected from above.

Next, upper message 7. Gently sink the upper electrode 5 to which -7 is attached. When the liquid 10 passes through the lower electrode 6 during injection, it accumulates a little towards the bottom, and on the other hand, when the electrode 5 is submerged, the liquid 10 flows upward.
O goes out and is captured by #IIO2 top and bottom mesh, 8.

With the cells 9 in a stacked state, that is, with the desired cells adhered to each other, the tube 11 is put in, the lid 2 is closed, and direct current pulses are applied from the upper and lower terminals 3.4 to effect a predetermined cell electrical fusion.

(g) Effects According to the present invention, cell fusion can be performed using only a DC bus generator without using a centrifuge or an AC power source.

The device is therefore very inexpensive.

[Brief explanation of the drawing]

FIG. 1 shows a cell fusion chamber according to the present invention, and FIG. 2 shows a cell fusion chamber using a centrifuge. 5-upper [pole 6-lower pole 7-upper mesh 8-・lower mesh 1: i: seal;
:

Claims (1)

[Claims] 1. A cell comprising a lower electrode provided at the bottom of a chamber body and an upper electrode that has a hole through which a cell suspension can pass and can be moved to a state where it can be brought into pressure contact with the lower electrode. fusion chamber. 2. The cell fusion chamber according to claim 1, wherein at least one of the lower electrode and the upper electrode is made of a conductive metal material having an insulating mesh and fine holes.