JPH08242885A - Measurement of activity of cell - Google Patents
Measurement of activity of cellInfo
- Publication number
- JPH08242885A JPH08242885A JP7047536A JP4753695A JPH08242885A JP H08242885 A JPH08242885 A JP H08242885A JP 7047536 A JP7047536 A JP 7047536A JP 4753695 A JP4753695 A JP 4753695A JP H08242885 A JPH08242885 A JP H08242885A
- Authority
- JP
- Japan
- Prior art keywords
- activity
- quinone
- cell
- hydrogen peroxide
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000000694 effects Effects 0.000 title claims abstract description 36
- 238000005259 measurement Methods 0.000 title description 10
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 claims abstract description 47
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 36
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims abstract description 34
- MJVAVZPDRWSRRC-UHFFFAOYSA-N Menadione Chemical compound C1=CC=C2C(=O)C(C)=CC(=O)C2=C1 MJVAVZPDRWSRRC-UHFFFAOYSA-N 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 17
- 229910052697 platinum Inorganic materials 0.000 claims abstract description 17
- 235000012711 vitamin K3 Nutrition 0.000 claims abstract description 14
- 239000011652 vitamin K3 Substances 0.000 claims abstract description 14
- 229940041603 vitamin k 3 Drugs 0.000 claims abstract description 14
- 229930192627 Naphthoquinone Natural products 0.000 claims 1
- 150000002791 naphthoquinones Chemical class 0.000 claims 1
- ZJTLZYDQJHKRMQ-UHFFFAOYSA-N menadiol Chemical compound C1=CC=CC2=C(O)C(C)=CC(O)=C21 ZJTLZYDQJHKRMQ-UHFFFAOYSA-N 0.000 abstract 1
- 229940100434 menadiol Drugs 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 39
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 20
- 239000006285 cell suspension Substances 0.000 description 11
- 235000019441 ethanol Nutrition 0.000 description 10
- 229910021607 Silver chloride Inorganic materials 0.000 description 6
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 6
- -1 phosphate compound Chemical class 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000004020 luminiscence type Methods 0.000 description 5
- 239000004332 silver Substances 0.000 description 5
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 229950006238 nadide Drugs 0.000 description 4
- 229910052709 silver Inorganic materials 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 230000037149 energy metabolism Effects 0.000 description 2
- 210000005260 human cell Anatomy 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 101710157142 2-methylene-furan-3-one reductase Proteins 0.000 description 1
- ACFIXJIJDZMPPO-NNYOXOHSSA-N NADPH Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](OP(O)(O)=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-N 0.000 description 1
- 101710189291 Quinone oxidoreductase Proteins 0.000 description 1
- 102100034576 Quinone oxidoreductase Human genes 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000006567 cellular energy metabolism Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000006056 electrooxidation reaction Methods 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、生細胞の活性を測定す
る方法に関する。TECHNICAL FIELD The present invention relates to a method for measuring the activity of living cells.
【０００２】[0002]
【従来の技術】新薬などを含む化学物質の安全性評価に
は、しばしば動物実験が行われているが、動物実験には
多大な費用と時間を要するため、近年動物実験代替法に
関する研究が数多くなされている。このような動物実験
代替法のひとつとして、生細胞を含む溶液に酸化型キノ
ンを添加し、溶液中に蓄積した過酸化水素を発光反応を
利用して検出・定量し、細胞内のエネルギー代謝活性を
測定する細胞活性測定方法が、山庄司らによる特開平１
−１６０４９９号公報、特開平２−２７６５９４号公
報、特開平３−２２８６９７号公報、特開平３−２６６
９９８号公報、特開平５−２９２９９８号公報に提案さ
れている。これらの公報で開示された方法の原理は、生
細胞内に存在するニコチンアミドアデニンジヌクレオチ
ド（ＮＡＤＨ）もしくはそのリン酸化合物（ＮＡＤＰ
Ｈ）を一方の基質として、酸化還元酵素ＮＡＤＨ（Ｐ）
Ｈ：キノン酸化還元酵素により生細胞を含む溶液に添加
されたキノン化合物が還元されて生じる還元型キノン化
合物がさらに溶液中の溶存酸素と反応して生じる過酸化
水素の溶液中蓄積量を発光反応を利用して測定するもの
である。2. Description of the Related Art Animal experiments are often conducted to evaluate the safety of chemical substances including new drugs, but since animal experiments require a great deal of expense and time, many studies on alternative methods for animal experiments have been conducted in recent years. Has been done. As one of the alternative methods for such animal experiments, oxidized quinone is added to a solution containing living cells, and hydrogen peroxide accumulated in the solution is detected and quantified using a luminescence reaction to measure intracellular energy metabolism activity. A method for measuring cell activity for measuring the activity is disclosed by Yamasho Tsukasa et al.
-160499, JP-A-2-276594, JP-A-3-228697, JP-A-3-266
It is proposed in Japanese Patent Laid-Open No. 998 and Japanese Patent Laid-Open No. 5-292998. The principle of the method disclosed in these publications is that nicotinamide adenine dinucleotide (NADH) or its phosphate compound (NADP) existing in living cells is used.
H) as one substrate, the oxidoreductase NADH (P)
H: Luminescence reaction of the accumulated amount of hydrogen peroxide in the solution, which is generated by the reaction of the reduced quinone compound produced by the reduction of the quinone compound added to the solution containing living cells by the quinone oxidoreductase, with the dissolved oxygen in the solution. Is used for measurement.
【０００３】[0003]
【発明が解決しようとする課題】従来の技術の項で述べ
た細胞活性測定では、過酸化水素を定量するために過酸
化水素と反応して発光をもたらす発光試薬が必要である
が、発光試薬が高価でかつ化学的に不安定であるという
問題があった。また、過酸化水素と発光試薬が反応して
生じる発光が不安定であるため、発光試薬を添加してか
ら発光量を測定するまでの時間を厳密に制御する必要が
あり、このような制御を行うために高価な自動測定装置
を利用するか、あるいは、測定操作に熟練した技術者を
養成する必要があった。In the cell activity measurement described in the section of the prior art, a luminescent reagent that reacts with hydrogen peroxide to generate luminescence is required to quantify hydrogen peroxide. Was expensive and chemically unstable. Further, since the luminescence generated by the reaction between hydrogen peroxide and the luminescent reagent is unstable, it is necessary to strictly control the time from the addition of the luminescent reagent to the measurement of the luminescence amount. It was necessary to use an expensive automatic measuring device to carry out or to train a technician skilled in the measurement operation.
【０００４】本発明の目的は、上述のような問題点を解
決し、簡便に生細胞の活性を測定できる方法を提供する
ことにある。An object of the present invention is to solve the above-mentioned problems and to provide a method for easily measuring the activity of living cells.
【０００５】[0005]
【課題を解決するための手段】本発明は、生細胞を含む
溶液に酸化型キノンを添加し、溶液中に生成する還元型
キノン又は過酸化水素を定量して前記細胞の活性を測定
する方法であって、生成する還元型キノン又は過酸化水
素を白金電極により検出、定量することを特徴とする。The present invention is a method for measuring the activity of cells by adding oxidized quinone to a solution containing living cells and quantifying reduced quinone or hydrogen peroxide produced in the solution. The reduced quinone or hydrogen peroxide produced is detected and quantified by a platinum electrode.
【０００６】酸化型キノンとしては、例えば２−メチル
−１，４−ナフトキノン（メナジオン）などを用いるこ
とができる。As the oxidized quinone, for example, 2-methyl-1,4-naphthoquinone (menadione) can be used.
【０００７】[0007]
【作用】本発明の細胞活性測定方法は、生細胞を含む溶
液に酸化型キノンを添加し、溶液中に生成した還元型キ
ノンあるいは過酸化水素を白金電極により検出、定量し
て細胞活性を測定するものである。The cell activity measuring method of the present invention measures the cell activity by adding oxidized quinone to a solution containing live cells and detecting and quantifying reduced quinone or hydrogen peroxide produced in the solution with a platinum electrode. To do.
【０００８】生細胞では、細胞のエネルギー代謝によっ
て生じるニコチンアミドアデニンジヌクレオチド（ＮＡ
ＤＨ）及びそのリン酸化合物（ＮＡＤＰＨ）と、主とし
て細胞膜内に存在するＮＡＤ（Ｐ）Ｈ−キノン酸化還元
酵素の存在が知られている。また、特開平１−１６０４
９９号公報では、細胞培養溶液に添加したメナジオンを
始めとするキノン化合物が生細胞と反応して溶液中に過
酸化水素が蓄積されるという報告がある。本願発明者
は、この過酸化水素あるいは過酸化水素が生成される前
の反応で生成される還元型キノンが、細胞培養液中で白
金電極を用い検出可能であることを見いだし本発明の創
出に至った。In living cells, nicotinamide adenine dinucleotide (NA) produced by cellular energy metabolism
It is known that DH) and its phosphate compound (NADPH) and NAD (P) H-quinone oxidoreductase mainly present in the cell membrane. In addition, Japanese Patent Laid-Open No. 1-1604
In Japanese Patent Publication No. 99, there is a report that a quinone compound such as menadione added to a cell culture solution reacts with living cells to accumulate hydrogen peroxide in the solution. The present inventors have found that this hydrogen peroxide or reduced quinone produced in the reaction before hydrogen peroxide is produced can be detected using a platinum electrode in a cell culture solution, and to the creation of the present invention. I arrived.
【０００９】本発明の方法により、生細胞を含む溶液に
酸化型キノンを添加することにより溶液中に生成された
還元型キノンあるいは過酸化水素を白金電極により定電
位電解酸化を行って、電流測定により溶液中の還元型キ
ノンあるいは過酸化水素を定量することにより、細胞活
性を測定することができる。According to the method of the present invention, reduced quinone or hydrogen peroxide produced in a solution containing living cells by adding oxidized quinone is subjected to potentiostatic electrooxidation with a platinum electrode to measure current. The cell activity can be measured by quantifying reduced quinone or hydrogen peroxide in the solution.
【００１０】[0010]
【実施例】次に本発明の一実施例について図面を参照し
て説明する。第１の実施例は、酸化型キノンを添加し、
生成した還元型キノンを定量することによる細胞活性測
定方法に関する。大日本製薬株式会社製ヒト株化肝細胞
Ｈｅｐ Ｇ２を常法に従ってトリプシン処理後、遠心分
離により集め、細胞密度が１×１０7 ｃｅｌｌｓ／ｍｌ
になるようにＥａｇｌｅ ＭＥＭ培地に懸濁した。この
細胞懸濁液をそれぞれ２ｍｌずつ３枚の３５mmシャーレ
にとり、この内の２枚にはそれぞれ細胞活性を阻害する
エチルアルコールを細胞懸濁液に対し１０％、５％含有
するように添加し、１枚はエチルアルコールを無添加と
し対照用とした。これらを二酸化炭素濃度５％、飽和水
蒸気、３７℃の条件で１時間培養した後、図１に示した
ように測定用容器５に細胞懸濁液４をとり有効電極面積
１１．１mm2 の白金作用電極１、対極２、銀・塩化銀参
照電極３を挿入し、白金作用電極１に銀・塩化銀参照電
極３を基準としてポテンシオスタット６を用いて０．２
Ｖの電位をかけて、白金作用電極１に流れる電流を測定
し、記録計７に１秒ごとに記録した。引き続き電位をか
けながら２００ｍＭメナジオン溶液を２０μｌ添加し
た。メナジオン溶液添加後直ちに電流値は上昇し、３分
後ほぼ添加前の値に戻った。図２は本実施例による測定
方法で測定した電流値の経時変化を説明するための模式
図である。メナジオン溶液添加前の定常的電流値はメナ
ジオン溶液を矢印８の時点で添加した後、曲線９のよう
に一時的な上昇を示した後、メナジオン添加前の定常的
な電流値に戻る。メナジオン添加前の定常電流値を延長
した線１０と曲線９で囲まれる領域１１の面積を細胞活
性値とした。その結果、エチルアルコールを添加しなか
った対照で得られた細胞活性値を１００とするとエチル
アルコールを１０％含む細胞懸濁液では細胞活性値は３
２、エチルアルコールを５％含む細胞懸濁液では８７と
なった。このように細胞活性を阻害するエチルアルコー
ルを添加した試料では、細胞活性値が小さくなってお
り、本発明によれば容易に細胞活性を測定することがで
き、また化学物質の安全性評価も行うことができる。An embodiment of the present invention will be described with reference to the drawings. The first example is the addition of oxidized quinone,
The present invention relates to a method for measuring cell activity by quantifying the produced reduced quinone. Human cell line Hep G2 manufactured by Dainippon Pharmaceutical Co., Ltd. was treated with trypsin according to a conventional method and then collected by centrifugation to give a cell density of 1 × 10 7 cells / ml.
Was suspended in Eagle MEM medium. Each 2 ml of this cell suspension was placed in three 35 mm Petri dishes, and ethyl alcohol, which inhibits the cell activity, was added to each of the two 35 ml dishes so as to contain 10% and 5% of the cell suspension, One sheet was used as a control without adding ethyl alcohol. These carbon dioxide concentration of 5% saturated steam, after incubation for 1 hour under conditions of 37 ° C., taking the cell suspension 4 into the measuring vessel 5 as shown in FIG. 1 of the effective electrode area 11.1 mm 2 platinum The working electrode 1, the counter electrode 2, and the silver / silver chloride reference electrode 3 are inserted, and the platinum working electrode 1 is adjusted to 0.2 by using the potentiostat 6 with the silver / silver chloride reference electrode 3 as a reference.
By applying a potential of V, the current flowing through the platinum working electrode 1 was measured and recorded on the recorder 7 every one second. Subsequently, 20 μl of 200 mM menadione solution was added while applying a potential. Immediately after the addition of the menadione solution, the current value increased, and after 3 minutes, it returned to the value before the addition. FIG. 2 is a schematic diagram for explaining the change over time in the current value measured by the measuring method according to this example. The steady-state current value before the addition of the menadione solution shows a temporary rise as shown by the curve 9 after the addition of the menadione solution at the time point of the arrow 8, and then returns to the steady-state current value before the addition of menadione. The area of the region 11 surrounded by the line 10 and the curve 9 in which the steady-state current value before the addition of menadione was extended was taken as the cell activity value. As a result, when the cell activity value obtained in the control to which ethyl alcohol was not added was 100, the cell activity value was 3 in the cell suspension containing 10% ethyl alcohol.
2 and 87 in the cell suspension containing 5% ethyl alcohol. As described above, in the sample to which ethyl alcohol that inhibits the cell activity is added, the cell activity value is small. According to the present invention, the cell activity can be easily measured, and the safety of chemical substances is also evaluated. be able to.
【００１１】次に酸化型キノンを添加することにより生
成する過酸化水素を定量することによる細胞活性測定の
一例を示す。大日本製薬株式会社製ヒト株化肝細胞Ｈｅ
ｐＧ２を常法に従ってトリプシン処理後、遠心分離によ
り集め、細胞密度が１×１０7 ｃｅｌｌｓ／ｍｌになる
ようにＥａｇｌｅ ＭＥＭ培地に懸濁した。この細胞懸
濁液を前記活性測定と同様に、それぞれ２ｍｌずつ３枚
の３５mmシャーレにとり、２枚にはそれぞれエチルアル
コールを１０％，５％になるように添加し、１枚はエチ
ルアルコールを無添加とし対照用とした。これらを二酸
化炭素濃度５％、飽和水蒸気、３７℃の条件で１時間培
養した後、図１に示したように測定用容器５に細胞懸濁
液４をとり有効電極面積１１．１mm2 の白金作用電極
１、対極２、銀・塩化銀参照電極３を挿入し、白金作用
電極１に銀・塩化銀参照電極３を基準としてポテンシオ
スタット６を用いて０．７Ｖの電位をかけて、白金作用
電極１に流れる電流を測定し、記録計７に１秒ごとに記
録した。引き続き電位をかけながら２００ｍＭメナジオ
ン溶液を２０μｌ添加した。メナジオン溶液添加後電流
値は徐々に上昇し、１分後ほぼ一定の値で安定した。こ
の時の電流値とメナジオン溶液添加前の電流値との差を
細胞活性値とした。その結果、エチルアルコールを添加
しなかった対照で得られた細胞活性値を１００とすると
エチルアルコールを１０％含む細胞懸濁液では細胞活性
値は３６、エチルアルコールを５％含む細胞懸濁液では
８４となった。本実施例においても、第１の実施例と同
様の結果が得られた。Next, an example of measurement of cell activity by quantifying hydrogen peroxide produced by adding oxidized quinone will be described. Dainippon Pharmaceutical Co., Ltd. human cell line hepatocyte He
pG2 was treated with trypsin according to a conventional method, collected by centrifugation, and suspended in Eagle MEM medium so that the cell density was 1 × 10 7 cells / ml. In the same manner as in the above-mentioned activity measurement, 2 ml each of this cell suspension was put into three 35 mm Petri dishes, and ethyl alcohol was added to each of the two plates so as to be 10% and 5% respectively. It was added and used as a control. After culturing these for 1 hour under the conditions of carbon dioxide concentration of 5%, saturated steam and 37 ° C., the cell suspension 4 was placed in the measuring container 5 as shown in FIG. 1 and the effective electrode area of 11.1 mm 2 of platinum was used. A working electrode 1, a counter electrode 2, and a silver / silver chloride reference electrode 3 are inserted, and a platinum potential of 0.7 V is applied to the platinum working electrode 1 by using a potentiostat 6 with the silver / silver chloride reference electrode 3 as a reference. The current flowing through the working electrode 1 was measured and recorded on the recorder 7 every second. Subsequently, 20 μl of 200 mM menadione solution was added while applying a potential. The current value gradually increased after the addition of the menadione solution, and became stable at a constant value after 1 minute. The difference between the current value at this time and the current value before the addition of the menadione solution was defined as the cell activity value. As a result, when the cell activity value obtained in the control to which ethyl alcohol was not added was 100, the cell activity value was 36 in the cell suspension containing 10% ethyl alcohol and the cell activity value in the cell suspension containing 5% ethyl alcohol. It became 84. Also in this example, the same result as that of the first example was obtained.
【００１２】[0012]
【発明の効果】以上説明したとおり、本発明の細胞活性
測定方法は、細胞の培養液に酸化型キノンを添加し、生
成した還元型キノンあるいは過酸化水素を白金電極によ
り検出し、細胞内のエネルギー代謝を測定し、細胞の活
性を測定する。本発明によれば、細胞活性測定に特別な
発光試薬などを用いずに、簡便かつ短時間に細胞の活性
を測定できる。また、測定に用いる白金電極は微小化で
きるため、装置が小型で、また少量の細胞懸濁液での測
定が可能である。As described above, according to the method for measuring cell activity of the present invention, the oxidized quinone is added to the cell culture solution, and the produced reduced quinone or hydrogen peroxide is detected by the platinum electrode to detect the intracellular Energy metabolism is measured and cell activity is measured. According to the present invention, cell activity can be measured simply and in a short time without using a special luminescent reagent or the like for measuring cell activity. In addition, since the platinum electrode used for measurement can be miniaturized, the device is small and measurement can be performed with a small amount of cell suspension.
【００１３】また、本発明により化学物質の安全性評価
を簡便に行うことができる。Further, according to the present invention, the safety of chemical substances can be easily evaluated.
【図１】本発明に関する白金電極を用いて細胞活性を測
定する装置の構成図である。FIG. 1 is a block diagram of an apparatus for measuring cell activity using a platinum electrode according to the present invention.
【図２】本発明の細胞活性測定法により得られる電流の
経時変化を示す模式図である。FIG. 2 is a schematic diagram showing the change over time of the electric current obtained by the cell activity measuring method of the present invention.
１ 白金作用電極 ２ 対極 ３ 銀・塩化銀参照電極 ４ 細胞懸濁液 ５ 測定溶液 ６ ポテンシオスタット ７ 記録計 ８ メナジオン溶液添加時点 ９ 電流値の経時的変化 １０ 定常電流値 １１ 領域 1 Platinum working electrode 2 Counter electrode 3 Silver / silver chloride reference electrode 4 Cell suspension 5 Measurement solution 6 Potentiostat 7 Recorder 8 Time point of addition of menadione solution 9 Time-dependent change in current value 10 Steady current value 11 Area
Claims (2)
し、前記溶液中に生成する還元型キノン又は過酸化水素
を定量して前記細胞の活性を測定する方法であって、生
成する前記還元型キノン又は前記過酸化水素を白金電極
により検出、定量することを特徴とする細胞活性測定方
法。1. A method for measuring the activity of the cells by adding oxidized quinone to a solution containing living cells and quantifying reduced quinone or hydrogen peroxide produced in the solution. A method for measuring cell activity, which comprises detecting and quantifying reduced quinone or the hydrogen peroxide with a platinum electrode.
ナフトキノン（メナジオン）である請求項１記載の細胞
活性測定方法。2. The oxidized quinone is 2-methyl-1,4-
The method for measuring cell activity according to claim 1, which is naphthoquinone (menadione).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7047536A JPH08242885A (en) | 1995-03-07 | 1995-03-07 | Measurement of activity of cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP7047536A JPH08242885A (en) | 1995-03-07 | 1995-03-07 | Measurement of activity of cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH08242885A true JPH08242885A (en) | 1996-09-24 |
Family
ID=12777863
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP7047536A Pending JPH08242885A (en) | 1995-03-07 | 1995-03-07 | Measurement of activity of cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH08242885A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003072428A (en) * | 2001-09-06 | 2003-03-12 | Suzuki Motor Corp | Control panel for driver |
| JP2007248071A (en) * | 2006-03-13 | 2007-09-27 | Horiba Ltd | Microorganism detecting method and microorganism detecting device |
| JP2012518161A (en) * | 2009-02-17 | 2012-08-09 | シーメンス アクチエンゲゼルシヤフト | Diagnostic equipment |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5255691A (en) * | 1975-09-24 | 1977-05-07 | Yellow Springs Instr | Improved diaphragm for enzyme electrode |
| JPS61500930A (en) * | 1984-01-21 | 1986-05-08 | ザ ブリテイツシユ ピトロ−リアム コンパニ− ピ−．エル．シ−． | Detection of membrane-bound systems |
| JPH03228696A (en) * | 1990-01-31 | 1991-10-09 | King Jozo Kk | Method for determining number of living yeast cells |
-
1995
- 1995-03-07 JP JP7047536A patent/JPH08242885A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5255691A (en) * | 1975-09-24 | 1977-05-07 | Yellow Springs Instr | Improved diaphragm for enzyme electrode |
| JPS61500930A (en) * | 1984-01-21 | 1986-05-08 | ザ ブリテイツシユ ピトロ−リアム コンパニ− ピ−．エル．シ−． | Detection of membrane-bound systems |
| JPH03228696A (en) * | 1990-01-31 | 1991-10-09 | King Jozo Kk | Method for determining number of living yeast cells |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003072428A (en) * | 2001-09-06 | 2003-03-12 | Suzuki Motor Corp | Control panel for driver |
| JP2007248071A (en) * | 2006-03-13 | 2007-09-27 | Horiba Ltd | Microorganism detecting method and microorganism detecting device |
| JP2012518161A (en) * | 2009-02-17 | 2012-08-09 | シーメンス アクチエンゲゼルシヤフト | Diagnostic equipment |
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