本文で2-iが参照されているのは以下ね。
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To investigate the relationship among STAP cells, STAP stem cells, Fgf4-induced stem cells, ES cells and trophoblast stem cells, we performed genome-wide RNA-sequencing analysis (Fig. 2i for dendrogram; Extended Data Figs 3 and 4 for expression analyses of representative genes ; Supplementary Tables 2 and 3 for analysis conditions).
それはLetter Extended Data Fig.6dのリジェンドに説明がある。
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d, Cluster tree diagram from hierarchical clustering of global expression profiles. Red, AU P values. As this analysis included morula and blastocyst embryos from which only small amounts of RNA could be obtained, we used pre-amplification with the SMARTer Ultra Low RNA kit for Illumina Sequencing (Clontech Laboratories).
関税というのは国家主権でどこにどれだけ掛けるかというのは自由だ。互いの
合意でやってるだけで、ブロック経済の反省からGATTができただけで、嫌いな奴に
対して関税撤廃なんて何の法的義務もないよ。もともとシナなんて資本主義経済研の中には
居なくて、資本主義側はシナなんかなくても何も困らない。シナから輸入できなくなったら
自分たちで作るようになるだけだ。価格が高くなっても、その金は所得を通して
国内で回るだけからね。Tax シナ to Save American Jobs.だよ。
桂報告の結論は両者に共通するX染色体の欠失と末端重複逆位接続により、
GLS1=GOF-ESだというものだね。これって4月、8月に既に解析されているね。
10月のは何の解析なのか。
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The line subjected to WGS is indicated in parentheses in cases in which several sublines were established for one cell type. Other sublines were confirmed by PCR and sequencing.
WGSはWhole Genome Sequencingだから意味だけ言うなら全ゲノム解析という
意味だ。細かな意味の違いがあるか、桂報告の本文が間違ってるんだろうね。
只。SNPs解析は全ゲノム読まないとあんな表は作れないんじゃないかな。
それだとBCA報告が間違っているか、<NGSによる全ゲノム解析>と<WGS解析>は
違う意味ということになるね。意味が違うならNGSでやっているか、もっとすごいので
やってるかの違いかな。でも、BCA報告の注には Other sublines were
confirmed by PCR and sequencing. と書かれている。
太田さんの2005年論文は<Generation of Normal Progeny by Intracytoplasmic
Sperm Injection Following Grafting of Testicular Tissue from Cloned Mice
That Died Postnatally>という題で若山さんとの共著だ。ここに129B6F1マウスが出てくる。
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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.
太田さんの2008年論文は<Increasing the Cell Number of Host Tetraploid
Embryos Can Improve the Production of Mice Derived from Embryonic Stem
Cells>という題で、 Hiroshi Ohta Yuko Sakaide Kazuo Yamagata Teruhiko Wakayama の
4人の共著だ。その中に129B6F1G1がでてきて、この参照論文が2005年論文になっている。
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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. 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]).
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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.
細胞塊の中にはおよそ1000個の細胞があるが、その中に数個Oct4蛋白を発現している
細胞塊が、19%程度あるということね。
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Of cell aggregates derived from liver cells treated with ATP, 19% expressed the amount of Oct3/4 comparable to ES cells (Fig. 3c). These data suggest that some proportion of cells in the aggregates express pluripotency-associated genes at comparable levels to those of ES cells.
Since the cell aggregates consist of ~10 cells, such expression level indicated possible existence of the cell(s) expressing pluripotency-associated genes at the equivalent level to that in ES cells.
In the present study, we investigated the properties of cell aggregates obtained by culture of liver cells transiently treated with low-pH stimulus, which was performed by the group directed by the author. We initially followed the protocol described in the original paper[1] with the detail description in protocol exchange where HCl was applied to achieve low-pH condition. However, we merely obtained the cell aggregates expressing the pluripotency makers as described in this report even when it was combined with the culture in medium containing Fgf2, which was not described in the original protocol but subsequently suggested by the authors. 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). However, the frequency was very low; 5 × 10<to the power of 5> 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%. Moreover, the pluripotency of such cells was not confirmed by chimera formation assay, and they did not give rise to any stem cell lines. We thus conclude that such cell aggregates do not fulfill the definition for STAP cells proposed in the original studies.
これかい。
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Since the cell aggregates consist of ~10 cells, such expression level indicated possible existence of the cell(s) expressing pluripotency-associated genes at the equivalent level to that in ES cells.
(英文)
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. We found that some aggregates expressed a comparable amount—more than 10% of the expression level in ES cells—of pluripotency-associated genes, including Oct3/4. Since the cell aggregates consist of ~10 cells, such expression level indicated possible existence of the cell(s) expressing pluripotency-associated genes at the equivalent level to that in ES cells. Klf4 expression was detected in all samples, which may reflect its expression in liver cells, and thus serves as a positive control in this assay. Of cell aggregates derived from liver cells treated with ATP, 19% expressed the amount of Oct3/4 comparable to ES cells (Fig. 3c). These data suggest that some proportion of cells in the aggregates express pluripotency-associated genes at comparable levels to those of ES cells.