JPS6384478A - Cell-fusion chamber - Google Patents

Abstract

(57) [Abstract] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

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

) Prior Art When using electrical stimulation to fuse cells, a cell suspension is placed in a cell fusion chamber that has a pair of opposing electrodes;
An AC electric field is applied between the electrodes. This brings all the cells floating in the cell suspension into close contact with each other, and in this state DC'
[Apply field to fuse cells.

e1 Problems to be Solved by the Invention Cells fused in a conventional chamber were randomly scattered within the chamber. Therefore, it is necessary to collect the fused cells using a cell acquisition device such as a micromanipulator or the like to remove the fused cells from the chamber and transfer them to a culture device.

Furthermore, in this case, it took a great deal of effort and time to collect all the cells, and great care was required to avoid damaging the cells.

This has been a bottleneck in culturing fused cells.

(2) Means for Solving All Problems The present invention is configured such that a pair of electrodes facing each other are made sufficiently long and thin with respect to the chamber space so that the density gradient of electric lines of force becomes large near the electrodes, and the electric A diaphragm is installed where the density gradient of the lines of force is large.

The diaphragm is composed of a 7 μlose membrane to which all cells are easily attached, or a synthetic fiber membrane treated to increase affinity with cells.

(g) Function The present invention is...! A diaphragm is located between the electrodes where a density gradient of air force lines is sufficiently generated, and fusion occurs on the diaphragm when cells are fully opened and brought together on the diaphragm. The septum to which the fused cells are attached is then removed from all chambers.

(f) Examples The present invention will be explained based on Examples. FIG. 1(a) is a schematic diagram of a chamber according to the present invention, and FIG. 1(bJ is a sectional view taken along the line AA in FIG. 1(a).

The fusion chamber C is provided with an electrode 1 and an electrode 2 corresponding to each other in a chamber body 4, and a connecting terminal 5 comes out from each electrode. The chamber main electrode 1.2 is sufficiently elongated with respect to the chamber space so that the density gradient of the electric lines of force becomes large in the vicinity of the electrode. A wire electrode can be considered as such an electrode. Then, near the electrode 1.2, where the density gradient of the lines of electric force becomes large, a heat-conducting diaphragm 3, 3', which has all the pores smaller than the diameter of the a-cell, is removably installed. A generator (not shown) applies an AC electric field between all electrodes. When an AC electric field is applied, the electric force is full! Since the maximum is near the electrode 1.2, the cells are attracted to the side with higher density, that is, the electrode. Cells drawn towards the electrode 1.2 attach here due to the presence of the septum 3.3'.

Fixed. A state diagram at this time is shown in FIG.

7 is a cell, and 8 is an electric field line.

In this way, for a while (with the ACC electric field generator, cells adhere to the cells and cause bearing). Even when bearing occurs, the DC-AC changeover switch (
DC pulses (e.g. 1,5 kV/Cm for plant cells,
30 μs).

When a DC pulse is applied to the cells, they cause fusion on the septum 3.3'.

When cells fuse on the septum 3.3', the septum 3.3'
is removed from chamber C and transferred to an incubation device (not shown).

In addition, in the above explanation, a method is described in which cells of different types A and B are suspended and fused together, but first, only the Am cells are placed in the entire chamber, and an AC electric field is applied to make the Ai cells adhere to the diaphragm. .

Then, when all the B cells have attached, the B cells are placed in the chamber. Then, B cells will adhere to A cells, and when D2 is applied in this state, A and B cells will fuse, and hybridomas (hybrid cells) can be efficiently produced.

(c) Effect Because the present invention fuses cells on the septum.

By transferring the entire septum to the culture device, it is unnecessary to pick up cells one by one using a micromanipulator.

In addition, since no mechanical manipulation is performed on the cells, the cells are not biased, which increases the subsequent growth activity.

Furthermore,! Cells that do not merge remain in the chamber when the septum is removed, increasing the probability of the number of fused cells among all the cells removed.

[Brief explanation of the drawing]

FIG. 1 (aJ is a schematic diagram of a chamber according to the present invention, FIG. 1(b) is a cross-sectional view of FIG. 1(a), and the second factor is an operational diagram of the present invention.

Claims (2)

[Claims] (1) In a cell fusion chamber that is equipped with a pair of electrodes facing each other and fuses cells by applying electrical stimulation to a cell suspension housed between these electrodes, the pair of electrodes is placed sufficiently within the chamber space. A cell fusion chamber characterized in that it is made elongated so that the density gradient of electric lines of force becomes large near the electrodes, and that a diaphragm is provided at a place where the density gradient of electric lines of force is large. (2) The cell fusion chamber according to claim 1, wherein the diaphragm is made of a cellulose membrane to which cells easily attach or a synthetic fiber membrane treated to increase affinity with cells.