JP3022106U - Cell fusion tank - Google Patents

Abstract

(57) Abstract: It is possible to increase the number of treatments performed under a stereomicroscope at a time and to keep the distance between electrodes constant in the longitudinal direction and the vertical direction of the electrodes, thereby exhibiting stable reproducibility. It is to provide a cell fusion tank capable of SOLUTION: Two or more grooves each having a rectangular cross-sectional shape are formed in parallel on the surface of a base, and a plurality of sets are formed in parallel. The electrodes were connected to the conductors of each set by protruding.

Description

[Detailed description of the device]

[0001]

[Technical field to which the device belongs]

 The present invention relates to a cell fusion tank, and more particularly to an electrical cell fusion tank that performs cell fusion by applying a voltage to cells.

[0002]

[Prior art]

 Nuclear transfer of mammalian eggs includes (1) egg micromanipulation technology, (2) artificial control of cell cycle, (3) electrical cell fusion technology, (4) parthenogenetic stimulation, (5) in vitro It consists of culture technology. Among these techniques, commercially available electric cell fusion devices are used for the electric cell fusion technique, and these commercially available devices have been developed to electrically fuse plant cells. Type is the mainstream, and nuclear transfer of mammalian eggs has been carried out by diverting the device body and its cell fusion tank.

The cell fusion tank is composed of a slide glass, a pair of wire electrodes arranged in parallel on the slide glass at predetermined intervals, and a petri dish containing the slide glass. The both ends of the wire electrode are linearly stretched on the upper surface of the slide glass by pressing the both ends with a pressing member. The fusion treatment by the cell fusion tank described above is performed by dropping the ovum between the wire electrodes with a pipette or the like under a stereomicroscope and applying the voltage required for the fusion to the wire electrodes.

[0004]

[Problems to be solved by the device]

 By the way, in the early stages of nuclear transfer research using animal ova, blastomeres at the 8-32 cell stage were used as donors in consideration of the totipotency of embryos, but recently, morula embryos and embryos have been used. The possibility of using inner cell mass of blastocyst or embryonic stem cell-like cells as donor blastomere has been reported. From these reports, it is inevitable that the fusion treatment will inevitably increase because the ratio of the size of the recipient egg to be fused and the size of the donor blastomere increases and the number of donor blastomere derived from one embryo increases. It is possible that the number will increase and the processing time will increase accordingly. However, in the conventional single-row parallel wire electrode fusion tank as described above, it is technically limited to handle 4 to 5 eggs at a time, and it is impossible to process a large number of eggs at one time. is there.

Further, as shown in FIG. 4 (a), the wire electrode 8 used in the above-mentioned fusion tank is a round wire having a circular cross-sectional shape, and therefore the intervals between the upper and lower parts of the wire are different. That is, the interval on the horizontal plane passing through the centers of both wires is the narrowest, and the interval becomes wider as going up and down with the horizontal plane as the boundary. As described above, the difference in the distance between the upper and lower parts of the wire electrode makes it difficult to perform cell fusion with good reproducibility because the electric field strength (stimulation condition) is different between the upper and lower parts. Furthermore, since the wire electrode 8 is only placed on the slide glass 9, it is easily moved by an external force, and if the distance between the wire electrodes changes, the reproducibility of cell fusion described above is greatly affected. . Moreover, there is a risk that the wire electrode may be displaced or peeled from the surface of the slide glass due to the force applied during cleaning and sterilization of the fusion tank, and the distance between the electrodes may be changed or damaged.

The present invention has been made in view of the problems of the above-described conventional technique. The purpose of the present invention is to increase the number of treatments under a stereoscopic microscope at one time and to reduce the distance between electrodes. It is an object of the present invention to provide a cell fusion tank that can hold the electrodes constantly in the longitudinal direction and the vertical direction and can exhibit stable reproducibility.

[0007]

[Means for Solving the Problems]

 The technical means taken by the present invention to achieve the above object is that a plurality of rows of reaction tanks are installed in parallel on a base made of a slide glass, and the conductors forming the reaction tanks have a rectangular cross section. A part of the conductor is fitted and fixed in a groove formed on the surface of the base. There are two or more rows of reaction tanks formed on the base, but when performing fusion processing under a stereoscopic microscope, set the rows so that they fit within one field of view (range that can be viewed simultaneously). In a preferred form, two rows of reaction tanks having a conductor interval of 1 mm are formed at 2 mm intervals.

With the above configuration, when fusion processing is performed under a stereoscopic microscope, a plurality of rows (two rows) of reaction tanks can be accommodated in one visual field, and a large number of eggs can be processed at one time. Since the conductors forming the reaction tank are fixed by partially fitting them into the grooves formed in the base, the intervals between the conductors do not change and are always kept constant. Furthermore, since the conductor has a rectangular cross-sectional shape, the space (distance) is constant anywhere in the upper and lower portions between the conductors, and the electric field strength is kept constant.

[0009]

[Embodiment of device]

 An embodiment of the present invention will be described below with reference to the drawings. In FIG. 1, reference numeral 1 denotes a base having a rectangular planar shape formed by a slide glass, and the surface of the base 1 has a rectangular sectional shape. The groove 2 is engraved at a predetermined interval, and a conductor 3 having a rectangular cross section having a width that allows the groove 2 to be tightly fitted is fitted in a part of the conductor 3 and fixed by protruding the other part onto the base 1. Has been done. One end of the conductor 3 in the longitudinal direction is crimped and fixed by an end plate 4 made of an insulating material (vinyl chloride resin), and the other is crimped and fixed by an electrode base 5 made of an insulating material (vinyl chloride resin) to which an electrode 6 is attached. At the same time, the electrode 6 is connected to the conductor 3.

In the groove 2 formed on the surface of the base 1, a pair of conductors 3 are arranged at a predetermined interval (for example, 1 mm), and such a set is parallel at a predetermined interval (for example, 1 mm to 2 mm). It is formed so as to be disposed in the. In the illustrated embodiment, the grooves are formed in a U-shape on a plane and arranged concentrically so that the electrodes do not have to be connected to each set, and the conductors 3 are fitted into the U-shape grooves. One of the electrodes is connected. Of course, the connection of the conductor and the electrode to the conductor is not limited to the illustrated form, and electrodes may be connected for each set.

The conductor fitted and fixed in the groove 2 is formed of a stainless wire having a rectangular cross section, and the conductor 3 has a lower half portion fitted and fixed in the groove 2 and an upper half portion from the surface of the base 1. It is installed so as to project upward. The conductor 3 fitted in the groove 2 is pressed by the end plates 4 and the electrode base 5 which are fixed to both sides of the base 1 with an adhesive, and is fixed so as not to be displaced or peeled off. Reference numeral 7 is a glass petri dish, and the base 1 having the conductor 3 attached thereto is housed in the petri dish 7.

[0012]

[Effect of device]

 The cell fusion tank of the present invention can perform more than twice the number of conventional treatments at one time due to the constitution of claim 1. As a result, even if the number of donor blastomeres increases, the fusion processing time will be the same as before or will be significantly shortened. In addition, since the conductors are fitted and fixed in the grooves formed in the base, the conductor spacing can be kept constant, and therefore the electric field strength can be kept constant at any position, and stable reproducibility is ensured. The conductor can be secured and the conductor is not displaced or peeled off from the base even when an external force is applied when cleaning and sterilizing the fusion tank, so that the cleaning and sterilization can be performed with peace of mind. Furthermore, since the conductors have a rectangular cross section, the vertical intervals between the conductors are the same, which makes it possible to stabilize the reproducibility described above.

[Brief description of drawings]

FIG. 1 is a partially cutaway plan view showing an embodiment of a cell fusion tank according to the present invention.

FIG. 2 is a front view of the same.

FIG. 3 is a sectional view taken along line (3)-(3) of FIG.

FIG. 4 is an explanatory view showing a conventional fusion tank and a fusion tank of the present invention.

[Explanation of symbols]

 1 ... Base 2 ... Groove 3 ... Conductor 6 ... Electrode

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Takahide Watanabe 3-19-16 Nukii, Nerima-ku, Tokyo Fujihira Industrial Co., Ltd. Fujimidai factory (72) Hideki Tsuchiya 3-Higashishinagawa, Shinagawa-ku, Tokyo 21-10 Livestock Improvement Corporation, Livestock Biotech Center

Claims (1)

[Scope of utility model registration request] 1. A plurality of sets are formed in parallel on the surface of a base housed in a petri dish with two grooves each having a rectangular sectional shape, and a part of a conductor having a rectangular section is fitted and fixed in the groove. A cell fusion tank, characterized in that the other part is projected onto the base and an electrode is connected to each set of conductors.