うん、そこはど素人なんでよくわからないな。相沢さんのマウスはどこかでてでくるかな?
肝臓で論文通りに近い数が出たんでそこを中心に調べてるな。
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Induced cell aggregates show poor induction of pluripotency-associated markers
To test the induction of pluripotency markers in cell aggregates obtained from ATP-treated liver cells, we assessed the expression of pluripotency-associated genes. Oct3/4 is a well-defined marker of pluripotent stem cells. Using a primer pair to detect Oct3/4 transcript from the Pou5f1 allele, but not pseudo-genes13, we did not find a detectable level (above 0.1% of the expression level in mouse ES cells, relative to the expression levels of Gapdh) of the transcript by quantitative polymerase chain reaction (Q-PCR) using a total RNA sample prepared from all cells in the culture (Fig. 3a), indicating that extremely few or no cells expressing Oct3/4 were present.
こんなことってあるのね。
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Interestingly, expression of Gfp from the Oct3/4-GFP transgene (GOF18)14 was detected in liver cells cultured for seven days irrespective of ATP treatment, suggesting leaky expression of this transgene.
そうだよ。丹羽さんも同じことをしている。
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We next performed qPCR on individual cell aggregates isolated from culture. Aggregates were selected and RNA samples were prepared separately. These RNAs were reverse-transcribed and qPCR was performed.
小保方さんはスフィア塊20個に1個Oct4発現があると報告したんだ。
それに対して、丹羽さんは同じ現象を再現確認したうえで、その細胞の数を数えた。
結果は
However, the frequency was very low; 5 × 105 liver cells yielded only ~30 cell aggregates, in which about 20% of the cell aggregates contained 1–2 Oct3/4 positive cells, indicating a frequency per seeded liver cell of 0.0012–0.0024%.
というものだ。105は10の5乗。
①から⑦までの現象ではこの細胞が多能性を持つということはできない。
小保方さんの論文の題は
<ティシュー誌>
The Potential of Stem Cells in Adult Tissues Representative of the Three Germ Layers
<博論>
Isolation of pluripotent adult stem cells discovered from tissues derived from all three germ layers
えっとね。ディスカッションのところでしょ。
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However, when we used ATP instead of HCl, also based on a suggestion by the authors, a few cells in a subset of cell aggregates expressed the pluripotency marker Oct3/4 at levels comparable to those in ES cells that were reproducibly detected by QPCR (Fig. 3c) and immunostaining (Fig. 4b).
ここと、
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Moreover, since the frequency of Oct3/4-positive cells in the cell aggregates was quite low, it was impossible to determine whether they were selected from the original population or induced in culture, again highlighting the lack of evidence supporting the existence of the reported STAP phenomenon.
だろ。
小町ちゃん、 丹羽さんが again highlighting the lack of evidence
supporting the existence of the reported STAP phenomenon. と言ってるのは
キメラができなかったことを理由にしているね。それから別途相沢さんが
この程度の数ではESですらテラトーマはできないと指摘している。違うかね。
<ティシュー誌>
The Potential of Stem Cells in Adult Tissues Representative of the Three Germ Layers
<博論>
Isolation of pluripotent adult stem cells discovered from tissues derived from all three germ layers