日本人は死を見ること帰するが如しだものね。いざとなったら座るのよね。
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4. Like most people, I am convinced that the mouse studies on STAP are solid and convincing. You personally have a top-notch reputation as a scientist around the world. People are instead more specifically concerned about the STAP stem cells themselves. One of the most common questions I am getting asked at this point is this–could the STAP cells have been contaminated with either mESCs or mouse iPS cells? Is that possible? How might that have happened?
まあ、機械のくせに人間様に意見するなんて。
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5. You mentioned that STAP stem cell is not working in your new lab. Why do you think that might be in terms of the methods? What is different now? Also, you mentioned that STAP worked for you when you were at RIKEN. Can you please be a bit more specific? Did you do 100% of the steps in the STAP induction yourself? Again, could iPS cells or ES cells somehow have gotten into the culture?
ふん、殺人機械は兵糧攻めに弱いのよね。そのときは電源切ってやる。
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Teru: I just learned one time from Dr. Obokata, then left RIKEN.
Do you know when we moved to a different lab in the past, how difficult it was then to reproduce even my own techniques? When I moved from Hawaii to Rockefeller, I spent half a year to reproduce cloned mice. This is my technique, but I still needed a lot of time. However, the method of STAP generation is not my technique, therefore, different lab and not my discovered techniques, so this is understandably more difficult to reproduce again.
I did 100% by myself, but each step was looked by Dr. Obokata. Much the same, one of my PhD student also succeeded to establish STAP-SC.
In those early stages of the experiment, we did not culture ES cell nor iPS cell at same time. After that, as control, we sometimes culture ES cell at same time.
サクサク行くんでしょ。メモしとこ。
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①When I succeeded to establish STAP-SC, the original STAP cells expressed Oct4-GFP very much.
②In addition, whole mRNA expression data suggest that STAP-SC are not ES cell.
③I did 100% by myself, but each step was looked by Dr. Obokata.
④Much the same, one of my PhD student also succeeded to establish STAP-SC.
⑤In those early stages of the experiment, we did not culture ES cell nor iPS cell at same time. After that, as control, we sometimes culture ES cell at same time.
ノフラーはSTAP細胞とSTAP幹細胞を論文に書かれているようには厳密に区別してないのよね。
だから彼がSTAP-SCと言ってるときSTAP-cellsと区別されてない。という以前に、論文を
信じてないから、STAP細胞をそのまま幹細胞認識しているのね。これは信じなければ当然の態度ね。
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6. Many people are trying the STAP method around the world and getting frustrated because it isn’t working. I know there is a methods paper in the works, but given how important this is would you consider putting a detailed step-by-step protocol out to the scientific community right now? For example, I’d be happy to post it on my blog and it would have no effect on the publication of a methods paper in a journal. This might really help people to get it to work. Waiting a month or two for a methods paper to come out might be too late.
I know there is a methods paper in the works, は丹羽さんのプロトコルの事じゃなくて
二報の論文の中に書かれているプロトコルを意味しているよね。このブログは2/27付で、
丹羽さんのプロトコル発表は3/5だ。ちなみに遠藤氏の「舐めてますね、これ」は3/9だ。
アーティクル、レターの両方にそれぞれのプロトコルが書かれている部分があるんだから
I know there are methods papers in the works, でないといけないわよね。ノフラーは
アーティクルのSTAPの作り方だけを意識しているのね。
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Teru: Yes, now RIKEN will soon publish the detailed method. I contributed chimera and to establishment. However, chimera and establishment is usual protocol, not specific, because it is the same or more easier than ES cell. Unfortunately, now all responsibility is in RIKEN, and I left RIKEN. Although I want to know but I do not know now.
はい。
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Teru: I do not escape. Because in my results everything is true. However, it is going to take time to reproduce the new technique. For example, the first cloned animal, Dolly, wasn’t reproduced for one and half years after it was published. Human cloned ES cell paper has still not yet been reproduced. Therefore, please wait at least one year. I believe that during this period someone or myself will success to reproduce it.
はい、やっと終りね。注よ。
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Note: At his request, I have made a scattering of small corrections to Teru’s answers just for a few typos/spelling/grammar issues since English is not his first language, but I did not change in any way the tone or significant content of his answers.
こんなものなしら。
①When I succeeded to establish STAP-SC, the original STAP cells expressed Oct4-GFP very much.
②In addition, whole mRNA expression data suggest that STAP-SC are not ES cell.
③I did 100% by myself, but each step was looked by Dr. Obokata.
④Much the same, one of my PhD student also succeeded to establish STAP-SC.
⑤In those early stages of the experiment, we did not culture ES cell nor iPS cell at same time. After that, as control, we sometimes culture ES cell at same time.
⑥However, chimera and establishment is usual protocol, not specific, because it is the same or more easier than ES cell.
⑦I doTherefore, please wait at least one year. I believe that during this period someone or myself will success to reproduce it.
⑧ I do not escape. Because in my results everything is true.
⑨since English is not his first language
検討の結果問題点は以下だったんだね。
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①When I succeeded to establish STAP-SC, the original STAP cells expressed Oct4-GFP very much.
②In addition, whole mRNA expression data suggest that STAP-SC are not ES cell.
③I did 100% by myself, but each step was looked by Dr. Obokata.
④Much the same, one of my PhD student also succeeded to establish STAP-SC.
⑤In those early stages of the experiment, we did not culture ES cell nor iPS cell at same time. After that, as control, we sometimes culture ES cell at same time.
⑥However, chimera and establishment is usual protocol, not specific, because it is the same or more easier than ES cell.
⑦I doTherefore, please wait at least one year. I believe that during this period someone or myself will success to reproduce it.
⑧ I do not escape. Because in my results everything is true.
⑨since English is not his first language
(2005年論文)
タイトル***Generation of Normal Progeny by Intracytoplasmic Sperm Injection Following Grafting of Testicular Tissue from Cloned Mice That Died Postnatally
URL***ttps://academic.oup.com/biolreprod/article/73/3/390/2666738
(2008年論文)
タイトル***Increasing the Cell Number of Host Tetraploid Embryos Can Improve the Production of Mice Derived from Embryonic Stem Cells
URL***ttps://academic.oup.com/biolreprod/article/79/3/486/2557580#60639901
何度でも繰り返そうかな。
2008年論文の実験は特許申請されているもので、この論文がベースになっている。
これは実在の実験だ。この論文のES Cell Linesのところに以下のようにある。
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The 129B6F1G1 [13] and BDmt2 [14] are nuclear transfer-derived ES (ntES) cell lines [15] previously established in our laboratory using Sertoli cells of 129B6F1 background with GFP and tail-tip cells of a male BDF1 mouse as donor for nuclear transfer, respectively.
この[13]の注が一番後ろにつけられているReferencesの13で2005年論文の事だ。
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13
Ohta H, Wakayama T. Generation of normal progeny by intracytoplasmic sperm injection following grafting of testicular tissue from cloned mice that died postnatally. Biol Reprod 2005; 73:390–395.
2005年論文には単に129B6という慣習表記で129が母親だとされているのではないのよ。
そんな間違いはないの。ちゃんと文章で母親が129だと書かれている。これほど念入り
なのもなかなかないわよ。2005年論文のMiceの項目よ。
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To generate 129B6F1 mice carrying the GFP transgene, female 129/Sv-ter mice were mated with male C57BL/6 GFP transgenic mice, and the offspring of these matings, which were hemizygous for the GFP transgene, were used as donors for nuclear transplantation.
female 129/Sv-ter mice と書かれてますよね。日経サイエンスに対して太田さんが
話していたTerがこれです。femaleっメスですよね。よはっきり書かれている。しかも、
実は2008年論文をさっき引用して13のリファレンスで2005論文のマウスが使われているのだと
指摘しましたが、その直後の文書は以下ですよ。
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Male 129B6F1 mice expressing GFP were generated by mating female 129/Sv mice with male C57BL/6 GFP transgenic mice (double-transgenic mouse line [16] constructed using pCAG-EGFP [10, 11] and Acr3-EGFP [17]).
ここはアルイミオウジさんも首をひねっていたところね。Ｔｅｒがないので省略と考えたようね。
ここはもう一度全文を貼りましょうね。
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ES Cell Lines
The D3 ES cell line was originally established by Doetschman et al. [12], and its GFP transgenic cell line (129SV(D3)-Tg(ACTB-EGFP)CZ-001-FM260Osb), which was established using pCAG-EGFP [10, 11], was kindly provided by Dr. Masaru Okabe (Osaka University, Osaka, Japan). The E14 ES cell line [7] was derived from the inbred mouse strain 129/Ola in 1985 by Dr. Martin Hooper in Edinburgh (Scotland) and was obtained through Dr. Peter Mombaerts (Rockefeller University). The 129B6F1G1 [13] and BDmt2 [14] are nuclear transfer-derived ES (ntES) cell lines [15] previously established in our laboratory using Sertoli cells of 129B6F1 background with GFP and tail-tip cells of a male BDF1 mouse as donor for nuclear transfer, respectively. Male 129B6F1 mice expressing GFP were generated by mating female 129/Sv mice with male C57BL/6 GFP transgenic mice (double-transgenic mouse line [16] constructed using pCAG-EGFP [10, 11] and Acr3-EGFP [17]).
以下はＥＳ細胞ではなくてマウスの説明だわよね。当然③を作るときに使われた核の説明になるわよね。ここで太田さんはこの核は雄だということを言ってる。
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Male 129B6F1 mice expressing GFP were generated by mating female 129/Sv mice with male C57BL/6 GFP transgenic mice (double-transgenic mouse line [16] constructed using pCAG-EGFP [10, 11] and Acr3-EGFP [17]).
2005年論文では雄雌を区別せずthe offspringとしている。これは集合名詞としての用法なので複数の子供ですね。常識で考えてもネズミが子供を一匹しか生まないなんてことはない。
2008年論文ではその中のオスから作られたntES細胞だと言ってることになる。
両親の雌雄は別として桂チームが分析したntESはオスだつた。オスだったからこそＸ染色体が母親からと決まっているから簡単に親の雌雄が違うと知れた。
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To generate 129B6F1 mice carrying the GFP transgene, female 129/Sv-ter mice were mated with male C57BL/6 GFP transgenic mice, and the offspring of these matings, which were hemizygous for the GFP transgene, were used as donors for nuclear transplantation.
でも親の雌雄はあくまでも129B6なのよね。太田さんは2005年論文は自分だけの
論文なのでこだわりがあるみたいね。2008年論文はいろんな人との共著だからね。
何度も両親が129Ｂ6だと繰り返するよね。そこは自分が作ったから詳しいから
ついつい詳しく書きたくなるのよね。問題はthe offspring of these matingsと
maitingが複数形になってるところよね。
まずないんだよ。Ooboeさん、アクロシンは精子で発現するんでメスでは光らない。
アクロシンの入っているのは岡部マウスだけど、そのメスを使うと普通のB6の
メスがコンタミすると子孫がAcr-GFPを持たなくなってしまう。確実にAcr-GFPを
持つ子供を作るにはどうしたらいいか。岡部マウスのオスから精子を取り出して
確実に光っている精子を使って複数のメスに顕微授精させればいいんです。
そうすると the offspring of these matingsは全て129B6になる。太田さんが
何度も確信をもってメスが129だと言ってるのはそれが理由だと思っていますよ。
ただ、そういうこまごまとしたことは追々はっきりしてくるはずです。何はともあれ、
論文がどうであるか以前に、ラベルと中身の違っているものを分析するなんて
どういう精神構造なのかと問えばいのです。
unknown abnormalitiesなのね。
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However, the usefulness of these techniques is limited by their low success rates; the birth of cloned offspring is quite rare, with average cloning efficiency rates of just 1–3%, regardless of the species [15]. In addition, cloned animals often die before sexual maturity as a result of unknown abnormalities [16].
