JP2002369677A - Conductive incubator and electrification system for the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a conductive incubator and an electrification system for the same for improving functions of a cultured cell, in vivo tissue, a microorganism by imparting an electrical stimulation, or imparting conductivity to the incubator for electrochemical measurement of the activities of these samples. SOLUTION: This invention modifies the surface of the incubator incubating the cell, in vivo tissue or the microorganism with a conductive material such as a metal, a metal compound, a cyclic compound, a polymer, carbon, and energizes them by suing the incubator itself as a working electrode, and putting in a counter electrode into a culture solution filled in the incubator and further energizing them by using the culture solution as an electrolyte while stably incubating the samples adhered on the surface of the incubator. The invention enables a long term energization to the cell, the in vivo tissue or the microorganism while stably culturing them, a constant long-term electrical stimulation to the sample under cultivation, and a long-term cultivation requiring a periodical electrochemical measurement.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a conductive incubator and an energizing system capable of energizing cultured cells, biological tissues, and microorganisms, so that these samples can be subjected to electrical stimulation by energization or the activity of these samples. This is effective when electrochemically measuring is used.

[0002]

2. Description of the Related Art Cultured cells, living tissues,
When energizing microorganisms or electrochemically measuring the activity of these samples, these samples have been forcibly adhered and cultured on a normal electrode surface. Of course, electrodes cannot be suitable for culturing cells, living tissues, microorganisms,
Practical use was extremely difficult when these samples were killed or contaminated with impurities in a short time.

[0003]

[Problems to be Solved by the Invention] It can be said that applying electrical stimulation to cultured cells, living tissues, and microorganisms to improve their functions and improve their viability is essential for the future development of the bioindustry. In addition, a technique for electrochemically measuring the activity of cultured cells, living tissues, and microorganisms is also an increasingly developed field in the future. However, until now, these samples have been forcibly adhered and cultured on the existing electrode surface, so there are many problems such as the death of these samples in a short period of time and contamination with impurities, making them difficult to use. Met.

[0004]

The gist of the technique for solving the problem is to make the incubator itself capable of stably culturing cells, biological tissues, and microorganisms conductive. Modification of the surface of containers for culturing cells, living tissues, and microorganisms with materials such as conductive metals, metal compounds, cyclic compounds, polymers, and carbon, and culturing cells, living tissues, and microorganisms in the incubator. Thereby, while stably culturing cells, living tissues, and microorganisms adhered to the surface of the incubator, it is possible to conduct electricity to these samples. The energization system uses the incubator itself as a working electrode, puts a symmetric electrode in a culture solution of cells, biological tissues, and microorganisms filled in the incubator, and energizes the culture solution as an electrolytic solution.

[0005]

The conductive incubator according to the present invention makes it possible for the first time to supply electricity to cultured cells, living tissues and microorganisms without disturbing these samples.
The present invention not only makes it possible to apply short-time electrical stimulation to cultured cells, living tissues, and microorganisms, but also makes it possible for the first time to constantly provide electrical stimulation during long-term culture. Furthermore, it has become possible for the first time to electrochemically measure the activity of these samples, not only once, as in the prior art, but periodically during culturing.

[0006]

EXAMPLES Hereinafter, a conductive incubator of the present invention and a current supply system thereof will be described based on examples. However, the examples do not limit the validity of this test.

[0007]

Embodiment 1 ITO (Indiu) having conductivity on the inner surface
FIG. 1 is a schematic view of an incubator on which an m Tin oxide) component is deposited and an energization system thereof. FIG. 2 shows a photograph of mouse hepatocytes cultured in this conductive culture vessel. As a result, the hepatocytes can be stably cultured in the ITO-evaporated conductive incubator, and the conductive incubator itself in which the hepatocytes are adherently cultured is used as a working electrode in a hepatocyte culture solution as an electrolytic solution. DC current is -0.
Load was applied in the voltage range of 5-0.5V. As a result, no change in the morphology of the hepatocytes due to energization was observed, and almost no stained hepatocytes were observed even when trypan blue for staining dead hepatocytes was added. FIG. 2 also shows a photograph of hepatocytes after energization and a photograph of hepatocytes stained with trypan blue after energization. From the above results, it was shown that the use of the conductive incubator according to the present invention allows stable supply of electricity to cultured hepatocytes.

[0008]

(1) According to the present invention, for the first time, it is possible to energize cells, living tissues, and microorganisms cultured in an incubator while using the incubator. It is preferable to use them when electrochemically measuring the activity of those samples or when applying an electric stimulus by energization without moving the sample to the same. (2) According to the present invention, after culturing cells, living tissues, and microorganisms for the first time without being contaminated by impurities, it becomes possible to energize these samples for a long period of time, and these samples are kept cultured. It is particularly preferable to provide a constant long-term electrical stimulation or to use for a long-term culture that requires periodic measurement of electrochemical activity. (3) The above procedure simplifies the operation process required to energize the cultured cells, living tissues, and microorganisms, resulting in significant cost reduction. (4) Long-term electrical stimulation of cultured cells, living tissues, and microorganisms became possible for the first time, and the function and viability of these samples could be improved by energization.

[0009]

[Changeable range] (a) As long as at least one substance capable of culturing cells, biological tissues, and microorganisms such as synthetic polymers such as glass and plastics, metals, and natural polymers is used in the incubator, Any material or form may be used. (B) As the conductive substance for modifying the incubator, any substance may be used as long as at least one conductive substance such as a metal, a metal compound, a polymer, a cyclic compound, and carbon is used. (C) The condition for modifying the incubator with a conductive substance may be any condition as long as at least cells, living tissues and microorganisms being cultured can be energized. (D) The culture solution used as the electrolytic solution may have any composition as long as it is at least energizable.

[0010]

[Impossible range] (I) In the current supply system, a conductive incubator is regarded as a working electrode, and there is at least one set of symmetrical electrodes with respect to the working electrode. (II) The conditions such as voltage, current, direct current alternating current, and frequency at the time of energization or electrochemical measurement may be any conditions and combinations as long as they can be used for cultured cells, living tissues, and microorganisms. (III) The cells, living tissues, and microorganisms to be cultured may be separated from the incubator at other times as long as they adhere to the incubator at least at the time of energization.

[0011]

[Usage] (A) It can be used as an incubator for culturing cells, living tissues, and microorganisms used for screening in combination with measurement of electrochemical activity. Since screening is a business, it is a right of the present invention. (I) The functions of cells, living tissues, and microorganisms can be controlled by electrical stimulation, and are the foundation of the bioindustry using cells, living tissues, and microorganisms.

[0012]

[Brief description of the drawings]

[FIG. 1] ITO (Indium) having conductivity on the inner surface
FIG. 2 is a schematic view of an incubator on which a Tin Oxide (Tin Oxide) component is vapor-deposited and a current supply system thereof.

FIG. 2 is a photograph of mouse hepatocytes cultured in a conductive incubator according to the present invention.

[0013]

[Explanation of symbols]

1 Incubator 2 Conductive substance coated on the surface of the incubator (ITO: I
ndium Tin Oxide) 3 Platinum wire of symmetrical electrode 4 Culture solution serving as electrolytic solution 5 Cultured hepatocytes

[0014]

[Explanation of symbols]

11 Conductive substance (ITO: Indium Tin O)
xide) Photograph of hepatocytes cultured on the surface of an incubator coated 12 Photograph of cultured hepatocytes after energization using a conductive incubator 13 Photograph of cultured hepatocytes after energization stained with trypan blue

 ──────────────────────────────────────────────────続 き Continuing from the front page (72) Yoshiyuki Kondo, Inventor 3-15-1, Tsuneda, Ueda-shi, Nagano Prefecture, Department of Functional Polymer Science, Faculty of Textile Science and Technology, Shinshu University (72) Inventor Toshihiro Akaike 3-1-1 Asahi, Matsumoto-shi, Nagano Prefecture No. 1 Shinshu University Graduate School of Medicine (72) Inventor Naoki Negishi 3-1-1 Asahi, Matsumoto City, Nagano Prefecture Shinshu University Graduate School of Medicine (72) Inventor Hirohiko Ise 3-1-1 Asahi, Matsumoto City, Nagano Prefecture No. 1 Shinshu University Graduate School of Medicine (72) Inventor Nobuhiro Sugihara 3-1-1 Asahi, Matsumoto City, Nagano Prefecture Shinshu University Graduate School of Medicine (72) Inventor Masaaki Kobayashi 3-chome Asahi, Matsumoto City, Nagano Prefecture No. 1 Shinshu University Graduate School of Medicine (72) Inventor Akira Tsukikawa 3-15-1, Tsuneda, Ueda-shi, Nagano Pref. (Reference) 4B029 AA01 CC01 DG10 EA20

Claims (2)

[Claims] 1. A conductive incubator characterized in that cells, living tissues and microorganisms can be cultured on the surface thereof, and electricity can be supplied while culturing cells, living tissues and microorganisms. 2. An energization system using a conductive incubator capable of energizing while culturing cells, living tissues, and microorganisms.