他研究者の論文からの文章剽窃（盗用） １件目

疑惑論文１：　Nature Article　や特許における文章剽窃（盗用）疑惑　1件目　について。（2件目はこちら）

小保方晴子らのNature Article論文の”Methods”のセクションにおける”Karyotype analysis”の文章の一部は、ドイツの研究者（Jianli Guoら）が2005年にIn Vitro Cell Dev Biol Anim.誌で発表した論文の文章から拝借したものですが、拝借元の論文を引用していないことが明らかとなり、問題となっています。
下記の文章で、赤く太文字でハイライトされている部分がJianli Guoらの論文と同一の文章です。 さらに、青く太文字でハイライトされている部分が、Jonathan Landryの博士論文（Dissertation）と同一の文章です。

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Guo Jianliらの論文との比較

赤く太文字でハイライトされている部分が同一

小保方氏らのNature Article論文：　"Stimulus-triggered fate conversion of somatic cells into pluripotency"

Nature 505, 641–647 (30 January 2014) 
著者：　Haruko Obokata　（小保方晴子）, Teruhiko Wakayama　（若山照彦）, Yoshiki Sasai　（笹井芳樹）, Koji Kojima　（小島宏司）, Martin P. Vacanti　（マーティン・バカンティ）, Hitoshi Niwa　（丹羽仁史）, Masayuki Yamato　（大和雅之）, Charles A. Vacanti　（チャールズ・ヴァカンティ）

Karyotype analysis

Karyotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAP stem cells were arrested in metaphase by colcemid (final concentration 0.270 µg ml−1) to the culture medium for 2.5 h at 37 °C in 5% CO2. Cells were washed with PBS, treated with trypsin and EDTA (EDTA), re-suspended into cell medium and centrifuged for 5 min at 1,200 r.p.m. To the cell pellet in 3 ml of PBS, 7 ml of a pre-warmed hypotonic 0.0375 M KC1 solution was added. Cells were incubated for 20 min at 37 °C. Cells were centrifuged for 5 min at 1,200 r.p.m. and the pellet was re-suspended in 3–5 ml of 0.0375 M KC1 solution. The cells were fixed with methanol/acetic acid (3:1; vol/vol) by gently pipetting. Fixation was performed four times before spreading the cells on glass slides. For the FISH procedure, mouse chromosome-specific painting probes were combinatorially labelled using seven different fluorochromes and hybridized as previously described⁴¹. For each cell line, 9–15 metaphase spreads were acquired by using a Leica DM RXA RF8 epifluorescence microscope (Leica Mikrosysteme GmbH) equipped with a Sensys CCD camera (Photometrics). Camera and microscope were controlled by the Leica Q-FISH software (Leica Microsystems). Metaphase spreads were processed on the basis of the Leica MCK software and presented as multicolour karyograms.

Guo Jらの論文：　Multicolor karyotype analyses of mouse embryonic stem cells.

• In Vitro Cell Dev Biol Anim. 2005 Sep-Oct;41(8-9):278-83.
• Guo J1, Jauch A, Heidi HG, Schoell B, Erz D, Schrank M, Janssen JW.
• 1Institute of Human Genetics, University of Heidelberg, D-69120 Heidelberg, Germany.

• Materials and Methods 
• Chromosome preparation
• Metaphase spreads of the ES cells were performed as follows. Subconfluent ES cells were arrested in metaphase by adding colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37° C in 5% CO₂. Cells were washed with PBS, treated with trypsin-ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml of a prewarmed hypotonic 0.0375 M KCl solution was added. Cells were incubated for 20 min at 37° C. Cells were centrifuged for 5 min at 1200 rpm and the pellet was resuspended in 3–5 ml of 0.0375 M KCl solution. The cells were fixed with methanol/acetic acid (3:1, vol:vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides.

• Multicolor FISH analysis (M-FISH)
  For M-FISH analysis mouse chromosome–specific painting probes were combinatorially labeled using seven different fluorochromes and hybridized as previously described (Jentsch et al., 2003). For each cell line 9–15 metaphase spreads were acquired by using a Leica DM RXA RF8 epifluorescence microscope (Leica Mikrosysteme GmbH, Bensheim, Germany) equipped with a Sensys CCD camera (Photometrics, Tucson, AZ). Camera and microscope were controlled by the Leica Q-FISH software (Leica Microsystems Imaging solutions, Cambridge, United Kingdom). Metaphase spreads were processed on the basis of the Leica MCK software and presented as multicolor karyograms.

なお、文章の拝借元における塩化カリウムを意味する"KCl"という正しい表記が、小保方論文では"KC1"という無意味な言葉になっています。さらに、小保方論文では、EDTA (EDTA)　と、ethylenediaminetetraacetic acidの略称であるEDTAを誤って連続して記載しています。なぜ、このような初歩的ミスが起こったかは不明ですが、自分で文章を作成せず、他者論文の文章を拝借したことが一因であるかもしれません。

上記の拝借された文章の中には、ライカ社（Leica Mikrosysteme GmbH）の DM RXA RF8 落射蛍光顕微鏡（epifluorescence microscope ）と、フォトメトリクス社 (Photometrics)の Sensys CCD カメラの実験機器名も含まれています。小保方氏らは、実験方法の文章を拝借するだけでなく、わざわざ、Guo Jianliらドイツの研究者が用いた実験機器と同一のものを用いて実験をしたのでしょうか？そのようなことが果たして可能なのでしょうか？

このライカの DM RXA 顕微鏡も、フォトメトリクスの Sensys CCD カメラも、1990年代末から2000年代前半に販売されていたもので既に製造中止になっている機種です。 また、Sensys カメラのウェブサイトには、 「Then turn your computer back on and boot Windows 98/2000/ME/XP again.」と記載されており、Win 98が現役だったような時代の懐かしい製品です。よって、小保方氏らが研究室を立ち上げるときに、このような古い実験機器を新規に購入することは不可能であり、また中古品も出回っていません。

(フォトメトリクスの パンフレットによると、このSensys CCD カメラ は専用PCIカードで画像を取り込むタイプで、 30～200万画素程度のものでカメラに放熱フィンが付いている非常に古いカメラです。)

小保方ら論文著者はKC1などのOCRの誤認識に基づく誤記をそのままにして剽窃していることから、ほとんど剽窃文章の中身を吟味していないことが推測され、実験機器名に関しても実際には使用していないにも関わらずそのまま剽窃して記載してしまった可能性があります。これらのことから、そもそも、このMaterials and Methodsに書かれているKaryotype analysis（核型分析）の実験は、論文の記述通りには行われていなかった可能性が推測されます。これらの疑惑を晴らすためには、小保方研究室、笹井研究室、若山研究室、ヴァカンティ研究室、あるいは理研などの彼らが所属する研究施設の共有実験機器のいずれかに上記の実験機器が存在し、これらの機器を使用して実際に実験を行ったことを生データにより示すしかありません。あるいは、論文内には外注したとは記載されていませんが、もし、この実験が外注により外部機関により行われていたとしたら、その外部機関から送られてきた報告書を確認する必要があるでしょう。

なお、Guo Jianli氏らの原論文では「DM RXA RF8 epifluorescence microscope (Leica Mikrosysteme GmbH, B ensheim, Germany) equipped with a Sensys CCD camera (Photometrics, Tucson, AZ). 」 と書いてあったのを、小保方氏らの論文ではわざわざメーカーの所在地や国名を削除して書き換えています。 もし、実際はもっと新しい機種や他のメーカーの機種を使っていたのなら、わざわざ実際使ったのとは違う機種の名前を残しつつ国名だけ消すのではなく、実際に使用している機種に合わせて記述を直すはずです。（執筆者ではなくてコピーエディターが書き換えた可能性も否定できませんが）。

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小保方氏らの特許：　さらに、この文章は、小保方晴子氏らが出願した特許（原文ＰＤＦ）中の下記文章の一部においても拝借されています。特許においても、KC1という誤記が見られます。さらに、Guo論文における"imaging"が、特許においては"hanging"という文脈上間違った単語に変換されています。

[00290] Karyotype analysis. Karyotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAPS cells were arrested in metaphase by colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37°C in 5% CO2. Cells were washed with PBS, treated with trypsin/ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml of a prewarmed hypotonic 0.0375 M KC1 solution was added. Cells were incubated for 20 min at 37°C. Cells were centrifuged for 5 min at 1200 rpm and the pellet was resuspended in 3-5 ml of 0.0375 M KC1 solution. The cells were fixed with methanol/acetic acid (3: 1; vol/vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides. For the FISH procedure, mouse chromosome-specific painting probes were combinatorially labeled using seven different fluorochromes and hybridized as previously described (Jentsch et al, 2003). For each cell line, 9-15 metaphase spreads were acquired by using a Leica DM RXA RF8 epifluorescence microscope (Leica Mikrosysteme GmbH, Bensheim, Germany) equipped with a Sensys CCD camera (Photometries, Tucson, AZ). Camera and microscope were controlled by the Leica Q-FISH software (Leica Microsystems hanging solutions, Cambridge, United Kingdom). Metaphase spreads were processed on the basis of the Leica MCK software and presented as multicolor karyograms.

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Jonathan Landryの博士論文（Dissertation）との比較

青く太文字で強調されている部分が同一　

小保方氏らのNature Article論文：　"Stimulus-triggered fate conversion of somatic cells into pluripotency"

Nature 505, 641–647 (30 January 2014) 
著者：　Haruko Obokata　（小保方晴子）, Teruhiko Wakayama　（若山照彦）, Yoshiki Sasai　（笹井芳樹）, Koji Kojima　（小島宏司）, Martin P. Vacanti　（マーティン・バカンティ）, Hitoshi Niwa　（丹羽仁史）, Masayuki Yamato　（大和雅之）, Charles A. Vacanti　（チャールズ・バカンティ）

Karyotype analysisKaryotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAP stem cells were arrested in metaphase by colcemid (final concentration 0.270 µg ml−1) to the culture medium for 2.5 h at 37 °C in 5% CO2. Cells were washed with PBS, treated with trypsin and EDTA (EDTA), re-suspended into cell medium and centrifuged for 5 min at 1,200 r.p.m. To the cell pellet in 3 ml of PBS, 7 ml of a pre-warmed hypotonic 0.0375 M KC1 solution was added. Cells were incubated for 20 min at 37 °C. Cells were centrifuged for 5 min at 1,200 r.p.m. and the pellet was re-suspended in 3–5 ml of 0.0375 M KC1 solution. The cells were fixed with methanol/acetic acid (3:1; vol/vol) by gently pipetting. Fixation was performed four times before spreading the cells on glass slides. For the FISH procedure, mouse chromosome-specific painting probes were combinatorially labelled using seven different fluorochromes and hybridized as previously described⁴¹. For each cell line, 9–15 metaphase spreads were acquired by using a Leica DM RXA RF8 epifluorescence microscope (Leica Mikrosysteme GmbH) equipped with a Sensys CCD camera (Photometrics). Camera and microscope were controlled by the Leica Q-FISH software (Leica Microsystems). Metaphase spreads were processed on the basis of the Leica MCK software and presented as multicolour karyograms.

Jonathan Landryの博士論文（Dissertation by Jonathan Landry）：

(submitted to the Combined Faculties for the Natural Sciences and for Mathematics of the Ruperto-Carola University of Heidelberg, Germany for the degree of Doctor of Natural Sciences)

The genomic and transcriptomic landscape of HeLa cells.

• Presented by Jonathan Landry born in Lyon, France Oral examination: 06/12/2012

• 3.4.4 Multicolor fluorescent in situ hybridization (M-FISH)
• Metaphase spreads of HeLa cells were performed as follows. Subconfluent HeLa cells were arrested in metaphase by adding colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37° C in 5% CO2. Cells were washed with PBS, treated with trypsin-ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml of a prewarmed hypotonic 0.0375 M KCl solution was added. Cells were incubated for 20 min at 37° C. Cells were centrifuged for 5 min at 1200 rpm and the pellet was resuspended in 3–5 ml of 0.0375 M KCl solution. The cells were fixed with methanol/acetic acid (3:1, vol:vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides. For M-FISH analysis, chromosome–specific painting probes were labeled using DOP-PCR amplified DNA using 7 different fluorochromes in a combinatorial manner and hybridized as previously described [130]. Twelve metaphase spreads were acquired by using a Leica DM RXA RF8 epifluorescence microscope (Leica Mikrosysteme GmbH, Bensheim, Germany) equipped with a Sensys CCD camera (Photometrics, Tucson, AZ). Camera and microscope were controlled by the Leica MCK-FISH software (Leica Microsystems Imaging solutions, Cambridge, United Kingdom). Metaphase spreads were processed on the basis of the Leica MCK software and presented as multicolor karyotypes.

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小保方氏らのNature Article論文、文章が類似していたGuo氏やLandry氏らの論文の、３つのいずれの論文も、下記のJentsch I氏らのCytogenet Genome Res.誌の論文を引用していましたので、Jentsch氏の論文と比較してみましたが、全く異なる文章でした。つまり、小保方氏らの論文は実験方法を開発したJentsch氏らの論文から文章をコピペしたのではなく、Jentsch氏らを参考にして実験を行ったGuo氏らが作成した文章を剽窃した可能性が高いと思われます。なぜ、Landry氏の博士論文がGuo氏の論文に類似しているかどうかについては不明です。

Cytogenet Genome Res. 2003;103(1-2):84-8.

Seven-fluorochrome mouse M-FISH for high-resolution analysis of interchromosomalrearrangements.

Jentsch I1, Geigl J, Klein CA, Speicher MR.

Institut für Humangenetik, Technische Universität München, München, Germany.

（以下検証用にMaterials and Methodsの部分だけを引用）

Materials and methods

 Preparation of mouse painting probes and of fluorochrome DNA pools The generation of mouse chromosome-specific painting probes (Rabbitts et al., 1995) and their amplification by “primary” and “secondary DOPPCR” was described previously (Jentsch et al., 2001). Similar to our human 7-Fluor M-FISH approach (Azofeifa et al., 2000) we generated a DNA pool for each fluorochrome according to the labeling scheme shown in Fig. 2. For example, the DEAC-pool consisted of painting probes for chromosomes 3, 5, 10, 11, 14, and Y; the FITC pool of chromosomes 3, 8, 13, 15, 19, and X, and so on. Each painting probe was carefully calibrated to have the same fluorescent intensity after FISH according to our previously described protocols (Eils et al., 1998). After PCR labeling, the length of the DNA was adjusted by DNase I digestion to a size of 300– 700 bp.

Preparation of the mouse M-FISH hybridization mix The hybridization mix was prepared by overnight ethanol precipitation of 6 Ìl of the DEAC-, 7 Ìl of the FITC-, 6 Ìl of the Cy3-, 2.5 Ìl of the Texas Red- (= Cy3.5), 6 Ìl of the Cy5-, 2.5 Ìl of the biotin- (= Cy5.5) and 3 Ìl of the digoxigenin- (= Cy7) labeled PCR product, together with 60 Ìl of mouse Cot-1 DNA and 20 Ìg salmon sperm DNA. After centrifugation at 13,000 rpm for 30 min at 4 ° C followed by a washing step with 70 % ethanol, the pellet was resuspended in the hybridization mix, consisting of 50 % formamide, 20% dextran sulfate, and 2× SSC.

Induction of chromosomal aberrations and preparation of mouse chromosomes The mouse chromosome preparations from mouse cell lines AG12 TUBO and TSA were done as described previously (Weaver et al., 1999) with some modifications. In brief, the spleen was isolated from mouse and transported in a tissue culture flask with RPMI/FBS. After homogenization with RPMI, the cells were incubated in a culture flask with RPMI, FBS, concanavalin A, LPS, ß-mercaptoethanol. After 48 h incubation, the cells were treated with colcemid and 0.075 M KCl and fixed in methanol/acetic acid.

 Hybridization, post-hybridization washes and detection of indirectly labeled probes The probe mixture was denatured for 7 min at 78 ° C and then preannealed for about 20 min at 37 ° C. The slides were denatured in 70 % formamide, 2× SSC for about 2.5 min at 72 ° C. After passage through an ethanol series on ice, the slides were air-dried and the hybridization mixture was added to the slide. The hybridization field was sealed with a cover slip and rubber cement, and the slides were incubated for two nights at 37 ° C.
 Following hybridization, the slides were washed in 4× SSC/Tween three times 5 min each at 42 ° C to remove the cover slip and then three times (5 min each) with 1× SSC at 60 ° C. The slides were blocked in 3% BSA in 4× SSC/Tween and incubated for 30 min at 37 ° C. After blocking, avidin Cy5.5 (1:100 in 4% SSC/Tween plus 1% BSA) and anti-digoxigenin-Cy7 (1:50 in 4× SSC/Tween plus 1% BSA) were added to the slides and incubated for at least 45 min in a moist chamber at 37 ° C. The slides were then washed three times (5 min each) in 4× SSC/Tween at 45 ° C, counterstained with DAPI (4),6-diamidino-2-phenylindole) and mounted in p-phenylenediamine dihydrochloride antifade solution.

 Epifluorescence microscopy Slides were visualized using a Leica DMRXA-RF8 epifluorescence microscope equipped with special filter blocks (Chroma Technology, Brattleboro, VT) as described (Eils et al., 1998). For image acquisition, a Sensys CCD camera (Photometrics) with a Kodak KAF 1400 chip was used. Both the camera and microscope were controlled with Leica Q-FISH software (Leica Microsystems Imaging Solutions, Cambridge, UK). Images were analyzed using the Leica MCK-Software package (Leica Microsystems Imaging Solutions, Cambridge, UK/Eils et al., 1998).

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理研の野依良治理事長からの戒めの言葉

なお、小保方晴子氏らが所属する理化学研究所の野依良治理事長はAdv. Synth. Catal.誌において、以下のように、研究・出版倫理と不正行為について論説しています。

研究・出版における不正行為には、明白なデータねつ造だけでなく、 別の実験で得たスペクトルデータを偽って載せる、反応収率を過大に 表現するなど細かいものもあります。また、論文の剽窃(plagiarism) にもさまざまな形態があり、参考にした文献を故意に引用しない、 他の著者の論文から表現の一部を借りながら自分のものとして発表 するなども含まれます。特に後者は、論文のintroductionで化合物や 反応の重要性を述べるなど誰が書いても似たような内容になるときに 起こりやすく、また英語を母国語としない研究者が他の著者のうまい 表現を借りたくなる気持ちは分かるが、あくまで許されることではないと 指摘します。

参考→　理研・野依理事長が研究・出版倫理と不正行為についてAdv. Synth. Catal.誌に寄稿（ワイリー・サイエンスカフェ）