何度でも貼り付けるわ。
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However, as Fgf4-induced stem cells lay between STAP stem cells and trophoblast stem cells in the dendrogram, the possibility of contamination of STAP stem cells in the Fgf4-induced stem-cell population cannot be ruled out. Previous studies have indicated that inner cell mass (ICM)-type pluripotent cells can be removed from culture by treating the culture with a JAK inhibitor <16>(Extended Data Fig. 5a, b). In contrast, the JAK inhibitor treatment had no substantial effect on Oct4-GFP expression in Fgf4-induced stem-cell culture (Extended Data Fig. 5c, d; see Extended Data Fig. 5e, f for control). Expression of neither pluripotency markers (Fig. 2j) nor trophoblast markers (Fig. 2k) was substantially affected, indicating that pluripotency marker expression is unlikely to reflect contaminating STAP stem cells (ICM-type). Consistent with this idea, Fgf4-induced stem cells that were strongly positive for the trophoblast marker Itga7 (a surface marker for trophoblasts but not ES cells) also expressed high levels of Oct4-GFP (Extended Data Fig. 5g).
STAP幹細胞のコンタミと書いてあるよな。ESのコンタミだとは書いてないね。
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However, as Fgf4-induced stem cells lay between STAP stem cells and trophoblast stem cells in the dendrogram, the possibility of contamination of STAP stem cells in the Fgf4-induced stem-cell population cannot be ruled out.
しかしながら、Fgf4誘導幹細胞は、樹状図においてSTAP幹細胞と栄養膜幹細胞との間に位置しているので、Fgf4誘導幹細胞集団におけるSTAP幹細胞の混入の可能性を排除できない。
(Extended Data Fig. 5a, b)はOct4-GFP挿入ESの培地にJAKiを添加した実験だね。
結果は添加後GFP蛍光が消える。つまり多能性を失う。先行する論文の論旨に従えば、
多能性を維持していたJAK(ヤーヌスキナーゼ)が分解されて細胞が分化段階に
入ったということだね。
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Previous studies have indicated that inner cell mass (ICM)-type pluripotent cells can be removed from culture by treating the culture with a JAK inhibitor <16>(Extended Data Fig. 5a, b).
先行する研究では、内部細胞塊（ICM)タイプの多能性細胞は、JAK阻害剤で培養を処理することにより培養物から除去することができることを示した（拡張データ図5a,b）。
本文は以下の部分かしらね。
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Expandable pluripotent cell lines from STAP cells•
STAP cells have a limited self-renewal capacity under the conditions used for establishment (Fig. 2g and Extended Data Figs 2e and 5a). However, in the context of the embryonic environment, a small fragment of a STAP cell cluster could grow even into a whole embryo (Fig. 4f). With this in mind, we next examined whether STAP cells have the potential to generate expandable pluripotent cell lines in vitro under certain conditions.
(続き)
STAP cells could not be efficiently maintained for additional passages in conventional LIF+FBS-containing medium or 2i medium20 (most STAP cells died in 2i medium within 7 days; Extended Data Fig. 8a). Notably, an adrenocorticotropic hormone (ACTH)+LIF-containing medium (hereafter called ACTH medium) known to facilitate clonal expansion of ES cells36 supported outgrowth of STAP cell colonies. When cultured in this medium on a MEF feeder or gelatin, a portion of STAP cell clusters started to grow (Fig. 5a, bottom; such outgrowth was typically found in 10–20% of wells in single cluster culture using 96-well plates and in >75% when 12 clusters were plated per well). These growing colonies looked similar to those of mouse ES cells and expressed a high level of Oct4-GFP.
(続き)
After culturing in ACTH medium for 7 days, this growing population of cells, unlike parental STAP cells, could be passaged as single cells (Fig. 5a, bottom, and Fig. 5b), grow in 2i medium (Extended Data Fig. 8a) and expand exponentially, up to at least 120 days of culture (Fig. 5c; no substantial chromosomal abnormality was seen; Extended Data Fig. 8b, c). Hereafter, we refer to the proliferative cells derived from STAP cells as STAP stem cells.
(続き)
STAP stem cells expressed protein and RNA markers for pluripotent cells (Fig. 5d, e), showed low DNA methylation levels at the Oct4 and Nanog loci (Extended Data Fig. 8d), and had a nuclear fine structure similar to that of ES cells (Extended Data Fig. 8e; few electron-dense areas corresponding to heterochromatin). In differentiation culture25, 26, 27, STAP stem cells generated ectodermal, mesodermal and endodermal derivatives in vitro (Fig. 5f–h and Extended Data Fig. 8f, g), including beating cardiac muscles (Supplementary Video 4), and formed teratomas in vivo (Fig.5i and Extended Data Fig. 8h; no teratocarcinomas, n = 40). After blastocyst injection, STAP stem cells efficiently contributed to chimaeric mice (Fig. 5j), in which germline transmission was seen (Extended Data Fig. 8i). Even in tetraploid complementation assays, injected STAP stem cells could generate mice capable of growing to adults and producing offspring (Fig. 5k, l; in all eight independent lines, Extended Data Fig. 8j).
(続き)
In addition to their expandability, we noticed at least two other differences between STAP stem cells and parental STAP cells. First, the expression of the ES cell marker protein Esrrβ, which was undetectable in STAP cells (Extended Data Fig. 5d, e), was clearly seen in STAP stem cells (Fig. 5e). Second, the presence of H3K27me3 foci, which was found in a substantial proportion of female STAP cells, was no longer observed in STAP stem cells (Extended Data Figs 5f and 8k). Thus, STAP cells have the potential to give rise to expandable cell lines that exhibit features similar to those of ES cells.
マテメソは以下だね。
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STAP stem-cell conversion culture
For establishment of STAP stem-cell lines, STAP cell clusters were transferred to ACTH-containing medium36 on MEF feeder cells (several clusters, up to a dozen clusters, per well of 96-well plates). Four to seven days later, the cells were subjected to the first passage using a conventional trypsin method, and suspended cells were plated in ES maintain medium containing 20% FBS. Subsequent passaging was performed at a split ratio of 1:10 every second day before they reached subconfluency. We tested the following three different genetic backgrounds of mice for STAP stem-cell establishment from STAP cell clusters, and observed reproducible data of establishment: C57BL/6 carrying Oct4-gfp (29 of 29), 129/Sv carrying Rosa26-gfp (2 of 2) and 129/Sv × C57BL/6 carrying cag-gfp (12 of 16). STAP stem cells with all these genetic backgrounds showed chimaera-forming activity.
For clonal analysis of STAP stem cells, single STAP stem cells were manually picked by a thin-glass pipette, and plated into 96-well plates at one cell per well. The clonal colonies were cultured in ES medium containing 20% FBS, and expanded for subsequent experiments.
Article Figure 6とExtended Data Figure 8のリジェンドは以下だ。
(Article Figure 6)
a, Growth of STAP stem cells carrying Oct4-gfp. Scale bar, 50 μm. b, Dissociation culture of STAP stem cells to form colonies. Scale bar, 100 μm. c, Robust growth of STAP stem cells in maintenance culture. Similar results were obtained with eight independent lines. In contrast, parental STAP cells decreased in number quickly. d, Immunostaining of STAP stem cells for pluripotency markers (red). Scale bar, 50 μm. e, qPCR analysis of pluripotency marker gene expression. f–h, In vitro differentiation assays into three-germ-layer derivatives.
f, Ectoderm: Rx+/Pax6+ (retinal epithelium; 83%, n = 6). g, Mesoderm: troponin-T+ (cardiac muscle; 50%, n = 6). h, Endoderm: Sox17+/E-cadherin+ (endodermal progenitors; 67%, n = 6). Scale bar, 50 μm. i, Teratoma formation assays. Formation of keratinized epidermis (ectoderm; left), cartilage (mesoderm; middle) and bronchial-like epithelium (endoderm; right) is shown. Scale bar, 100 μm. j, Blastocyst injection assays. These pictures of live animals were taken serially (asterisk indicates the same chimaeric pup). k, l, Tetraploid complementation assay. ‘All-GFP+’ pups were born (k) and germline transmission was observed (l).
(Extended Data Figure 8)
a, Compatibility of 2i conditions with STAP stem-cell derivation from STAP cells and STAP stem-cell maintenance. STAP stem cells could not be established directly from STAP cells in 2i + LIF medium (top). However, once established in ACTH medium, STAP stem cells were able to survive and expand in 2i + LIF medium. Scale bar, 100 μm. b, Q-band analysis (n = 4; all cell lines showed the normal karyotype). c, Multicolour FISH analysis (n = 8; all cell lines showed the normal karyotype) of STAP stem cells. d, Methylation status of the Oct4 and Nanog promoters. e, Electron microscope analysis of STAP stem cells. Scale bar, 1 μm. f, g, Beating cardiac muscle (mesoderm; 38%, n = 8). Red line indicates an analysed region for kymograph (g).
h, Clonability of STAP stem cells. Clonal expansion from single STAP stem cells was performed. Pluripotency of clonal cell lines was confirmed by teratoma formation assay, showing the formation of neuroectoderm (left), muscle tissue (middle) and bronchial-like epithelium (right). Scale bar, 100 μm. i, Production of chimaeric mice from STAP stem-cell lines using diploid embryos. *These STAP stem-cell lines were generated from independent STAP cell clusters. j, Production of mouse chimaeras from STAP stem-cell lines by the tetraploid complementation method. *These STAP stem-cell lines were generated from independent STAP cell clusters. k, No H3K27me3-dense foci are seen in female STAP stem cells (n = 50; the CD45+ cell is a positive control). Scale bar, 10 μm.
これだけのものの移管が行われていて、その後処理として小保方さんが論文同士の整合を取ってるんでしょうね。笹井さんはそこまでは忙しすぎてやってないんじゃないかしら。STAP cells have a limited self-renewal capacity under the conditions used for establishment (Fig. 2g and Extended Data Figs 2e and 5a). のExtended Data Figs 2e ってTCRの説明よね。そこ全部変ね。
(続き)小保方さんは論文では主にリンパ球を使ってSTAP細胞を作っています。TCR再構成の実験をするときだけはCD90でもう一度T細胞だけをFACS選別します。でも厳格な選別ではないから、丹羽さんがヘテロな集団だと言ってるんですね。
マウスの白血球のもともとの構成割合はB細胞:60%、T細胞:30%、マクロファージ:5%、その他、5%です。脾臓の構成細胞全体での細胞の大きさは5から10マイクロメーターです。
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David Aphkhazava
23.19
Size of mouse splenocytes?
Does anyone know what the size of a mouse splenocyte is?
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Joseph michael Cantor
University of California, San Diego
5-10 um on average, with resting B and T cells comprising the lower end of the range and macrophages on the upper end. Adult mouse spleens typically contain 1x10e8 white blood cells: 50-60% B cells, 30% T cells, 5% macrophages, and <5% others.
[連投5]
参考
>while a small number aggregates gave rise to colonies containing small cells with large nuclei, resembling the morphology of embryonic stem cells. However, most of these cells ceased proliferation at day 7 and gradually regressed. <Investigation of the cellular reprogramming phenomenon referred to as stimulus-triggered acquisition of pluripotency (STAP) Hitoshi Niwa>
最後のテラトーマに関する記述は以下ね。
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Notably, when cultured in LIF+FBS-containing medium for 4 days, Fgf4-induced stem cells underwent substantial changes in morphology and started to form ES-cell-like compact colonies with strong GFP signals (Fig. 3a). These cells showed expression of pluripotency makers, but not trophoblast markers (Fig. 3b and Extended Data Fig. 6a), and formed teratomas in mice (Extended Data Fig. 6b). These ES-like cells were generated from Fgf4-induced stem cells sorted for strong expression of the trophoblast marker Itga7, but rarely from Itga7-dim cells (Fig. 3c, d).
"Gene Expression Profiling Reveals a Novel Regulatory Role for Sox21 Protein in Mouse Trophoblast Stem Cell Differentiation"
"Gene Expression Profiling Reveals a Novel Regulatory Role for Sox21 in Mouse Trophoblast Stem Cell Differentiation"
"A Resource for the Transcriptional Signature of Bona Fide Trophoblast Stem Cells and Analysis of Their Embryonic Persistence"
2019/3/21(木) 午後 0:20[ Ts.Marker ]返信する
(小保方論文 Published online 29-Jan-14)
Stimulus-triggered fate conversion of somatic cells into pluripotency
Bidirectional developmental potential in reprogrammed cells with acquired pluripotency
(遠藤論文 published online: 21 SEP 2014)
Quality control method for RNA-seq using single nucleotide polymorphism allele frequency
(Moretto Zita M 論文 Epub 2015 Oct 21.)
Gene Expression Profiling Reveals a Novel Regulatory Role for Sox21 Protein in Mouse Trophoblast Stem Cell Differentiation.
(Georg Kuales 論文 Published online 2015 Dec 14. )
A Resource for the Transcriptional Signature of Bona Fide Trophoblast Stem Cells and Analysis of Their Embryonic Persistence
(小保方論文 Published online 29-Jan-14)
Stimulus-triggered fate conversion of somatic cells into pluripotency
Bidirectional developmental potential in reprogrammed cells with acquired pluripotency
(遠藤論文 published online: 21 SEP 2014)
Quality control method for RNA-seq using single nucleotide polymorphism allele frequency
(Moretto Zita M 論文 Epub 2015 Oct 21.)
Gene Expression Profiling Reveals a Novel Regulatory Role for Sox21 Protein in Mouse Trophoblast Stem Cell Differentiation.
(Georg Kuales 論文 Published online 2015 Dec 14. )
A Resource for the Transcriptional Signature of Bona Fide Trophoblast Stem Cells and Analysis of Their Embryonic Persistence
僕の書き込み以前に書き込まれていたね。JBC論文というのは
(Moretto Zita M 論文 Epub 2015 Oct 21.)
Gene Expression Profiling Reveals a Novel Regulatory Role for Sox21 Protein in Mouse Trophoblast Stem Cell Differentiation.
のことなんで僕の指摘していね時系列的には遠藤論文の根拠にはなってないよね。
現在Sox21がどう考えられているかは今問題にしていない。
ただ、L氏は当初から「SOX21がTS特異的マーカーとは考えていません。」ということだったのかな。
新しい論文紹介だね。追加しておこう。L氏のスピン書き込みの根拠論文を
自分も読んでるという紹介でもあるんだね。
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(MubeenGoolam論文 24 March 2016)
Heterogeneity in Oct4 and Sox2 Targets Biases Cell Fate in 4-Cell Mouse Embryos
あなたの固定トゥイートは以下です。
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Coexpression of ES and TS markers can be seen in registered data of FI-SC Chip-seq which was regarded as ES.
How can we explain this fact?
そこの理解に関して一致できれば、今度は丹羽論文の曖昧さに関しても指摘できることに
なると思います。ES10個分の発現量と比較して0.1は1個分ですけど、これは一個だとは
言えませんよね。1/100が10個集まったら1/10です。1/1000が100個集まっても1/10です。
とても微量な発現がたくさんあるという可能性を否定できていませんね。（図4a）に2個の
スフィア塊の中で11個の個別細胞が免染で黄色く着色されているのが見えます。結論には
6個のスフィア塊の中に1から2個だったはずですね。これは1から2個分の発現と読み替え
ないといけないんでしょうね。
>>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%.
ティシュー論文の記述ね。
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Individual spheres possessed >2000 cells.
(The Potential of Stem Cells in Adult Tissues Representative of the Three Germ Layers:Sept.2010)
ティシュー論文に「Individual spheres possessed >2000 cells.(The Potential of Stem Cells in Adult Tissues Representative of the Three Germ Layers:Sept.2010)」とあります。STAP論文にもどこかにあったと思いますけど今見つけられません。いずれにせよ、勘違いなさっておられるので、図のブライトフィールドの三番目のスフィア塊の直径上に並んでいる細胞数を数えてみてください。12個程度でしょ。半径が6個のスフィアの体積は4πr^3/3で959個程度と逆残できます。写真はパラフィン固定して薄くスライスした中央付近の断面です。上は桑実胚なので16細胞期ですが、下は胚じゃなくてスフィア塊です。若山さんがナイフで切り分けてインジェクトするあれです。以上です。
The Potential of Stem Cells in Adult Tissues Representative of the Three Germ Layers
*ttps://www.researchgate.net/profile/Karen_Westerman/publication/47154811_The_Potential_of_Stem_Cells_in_Adult_Tissues_Representative_of_the_Three_Germ_Layers/links/556637fc08aefcb861d19869/The-Potential-of-Stem-Cells-in-Adult-Tissues-Representative-of-the-Three-Germ-Layers.pdf
[連投3]
まず①に関しては、これを主張しているのは桂報告書で<当事者は>調査チームであって、決して<沈黙して>いません。ご承知の通りBCA報告に至ってはタイトルがSTAP cells are derived from ES cellsとそのものです。調査チームの調べた細胞同士の関係は以下であったと主張しています。
①FLS→FES1(太田さんが実験にも使用せず作ってみたとされている岡部マウスとのF1由来受精卵ES細胞)
②GLS→GOF-ES(学生が小保方さんに渡したntES)
③CTS→FES1(太田さんが実験にも使用せず作ってみたとされている岡部マウスとのF1由来受精卵ES細胞)
遠藤さんの論文記載は以下だ。
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the FI‐SC population originated from two cell types: ESC‐like cells having a B6 genetic background and TSC‐like cells having a genotype similar to that of CD1
[連投1]
>>学さん
素晴らしい場を提供していただいて本当に感謝します。まず確認しなければならないことは、この実験は若山研で行われたものではないという事実ですね。Extended Data Figure 5-a,bの写真はOct4-GFPですね。小保方さんはOct4-GFPのESを若山研で入手はしていませんね。小保方さんが持っているのは学生が作って小保方さんがもらったと言われているOct4-GFPのntESです。さらにExtended Data Figure 5-e,fに使われているGFP/BFP共挿入マウスのESも小保方さんは若山研では入手していません。提供者は丹羽さん、もしくは笹井さんですが、ほぼ丹羽さんだと思われます。というのは公共データ登録されているTSはマウス背景がCD1と書かれていて、これは若山さんが作ったと言っている129B6F1CAGホモマウス背景のTSが分化していたため丹羽研から提供されたと報告されていますね。
[連投5]
Extended Data Figure3-bのヒートマップの下から1/3の辺りにPou5f1(Oct4)がありますね。CD45,ES,STAPの順に並んでいますが、右端のスケールで、CD45は2-4レヴェル、ESは10-12、STAPは4-6ですね。2を底とする対数表示ですから、リニアにはCD45は4-16、ESは1024-4096、STAPは16-64です。ESの0以上の発現遺伝子を真ん中に置いてESと同じ発現遺伝子に関してCD45とSTAPを比較している。これを見ると丹羽さんの後の検証確認と一致していますね。ESと比較したらSTAPは格段少ない。でも今はそのことを論じているのではなくて、Pou5f1(Oct4)はES特異的遺伝子であるが、Pou5f1(Oct4)が発現していたらESなのではないということの根拠として指摘しているんです。遠藤さんの論拠を思い出してください。TS特異的遺伝子が発現していたらTSだと言ってるんですよ。非論理も極まっている。
[連投8]
話を戻します。Extended Data Figure 5-a,b,c,dはESのOct4はJAKiの添加で消えるがFI-SCは消えないというものです。私が錯誤という言葉を使ったことが学さんの誤解を生んでいるようですが、ここの部分の本文は以下ですよね。
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However, as Fgf4-induced stem cells lay between STAP stem cells and trophoblast stem cells in the dendrogram, the possibility of contamination of STAP stem cells in the Fgf4-induced stem-cell population cannot be ruled out.
[連投10]
論文の趣旨に沿って、Extended Data Figure 5-e,fを見てみましょう。ここはいろんな人が指摘しているところですが、FI-SCのOct4-GFPはJAKiの添加で消えませんでしたよね。e,fで使われているFI-SCは無論c,dで使われたものと同じですね。白の矢印の先にFI-SCがある。矢印はブライトフィールドでの同じ位置を示していますね。eの中段はOct4-GFP細胞ですからJAKi添加前も後も光ってないといけませんね。c,dと同じ結果でないといけない。でもどちらも光ってない。最下段はどうかというと、これはブルーのフィルターなんですからどちらも緑色蛍光は見えないのが当然です。ここは問題ない。
第二レフェリーのアドヴァイスね。
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The authors suggest that STAP cell-derived TS-like cells are distinct from embryo-derived TS cells, but the data for this are not decisive and could be attributed to persistent contamination with STAP cells or ES cells (which could expain the occasional Nanog positive cells). Presence of ES cells could be excluded by culture in the presenve of JAK inhibitor, and monitored by Oct4 and Nanog immunostaining. A global transcriptome anallysis could then be performed for comparison with TS cells. Data should also be provided on Fgf4 withdrawal from the TS-like cells - does this induce the expected differentiation into trophoblast giant cells?
貼ってきたよ。
>>
[連絡1]
>>学さん
一日の連投量に制約があります。お返事は遅れます。ご了解願う。JAKiに関するネイチャー第二レフェリーの査読文だけ先に貼っておきます。
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The authors suggest that STAP cell-derived TS-like cells are distinct from embryo-derived TS cells, but the data for this are not decisive and could be attributed to persistent contamination with STAP cells or ES cells (which could expain the occasional Nanog positive cells). Presence of ES cells could be excluded by culture in the presenve of JAK inhibitor, and monitored by Oct4 and Nanog immunostaining.
[連絡2]
(続き)A global transcriptome anallysis could then be performed for comparison with TS cells. Data should also be provided on Fgf4 withdrawal from the TS-like cells - does this induce the expected differentiation into trophoblast giant cells?
[連投4]
第二レフェリーはSTAP細胞かES細胞がコンタミしている可能性があると言ってますね。その理由として時折Nanogの出ているデータがあると指摘している。で、もしESのコンタミならJAKiで取り除けると言ってるんですね。 could be excludedと表現している。取り除くという意味ですね。<constitutively expressed BFP (the number of plated cells was one-tenth of that of plated Fgf4-induced stem cells)>も存在していてはいけません。ただし、査読者はcouldと言ってますね。できるんじゃないかと言ってる。
[連投8]
先行する論文は参照文献で示されているが、この論文がレフェリーの言っているように、<Presence of ES cells could be excluded by culture in the presence of JAK inhibitor, >と主張しているかどうかは、我々が読んでも別問題だと分かりますが、笹井さんならもっと分かっているでしょうね。でもリジェクトされない作文が必要ですね。でも、そもそもそんなことしていいんでしょうかね。
[連投9]
学さんが先へ先へ進まれていることは承知しています。学さんらしく直感的表現で、<STAP論文を世に出すことの危険性を、なぜ、一流の学者たちが予想しなかったのか？ >と問われた真意は分かっています。アーティクルのアブストの中に以下の文章があって、彼の内心の葛藤を漏らしたものと思っています。彼は万が一に備えて自分のことを告白しているんですよね。
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Thus, our findings indicate that epigenetic fate determination of mammalian cells can be markedly converted in a context-dependent manner by strong environmental cues.
お答え申し上げます。多分今からでは入手困難と思います。私は2014年の事件発生当初、慶応大の吉村教授のブログでこんな流出論文がネット上に出回ってるぞと紹介されたPDFファイルのコピーを保存しています。
これは流出させてはいけないものだが、それをPDFファイル付きで出回ってるぞと知らせる行為は自分も流出拡散に協力していることなので変だなとは思いながらも本物だということはすぐにわかりました。この吉村教授の名は後に遠藤論文の謝辞 に、<I would like to acknowledge Dr Akihiko Yoshimura, Keio University, who initially suggested on his website that NGS data could be evaluated by SNP analysis.>とあることで若山さん側の関係者だと分かりました。
[連投2]
第二レフェリーがcould be excludedとアドヴァイズした言葉を笹井さんはcan be removed と言い直していますね。これはやんわりとES細胞は死滅してexcludeされたわけではないよねとレフェリーに指摘しているのに対して小保方さんはリジェンドの中でeliminatedと笹井さんの苦労を徒労にしてしまっています。cludeというのはcloseの意味のラテン語根ですね。eはex、limiはlimitの意味のラテン語根limesで、excludeとeliminateはほぼ同義だと思いますが、removeは別の場所に動かすことで、ESが死滅したというニュアンスはないですね。
[連投3]
先行論文はヤーヌスキナーゼが多能性を維持しているようだからその阻害剤を入れたら多能性細胞は分化し始めると言ってるんですね。ESであった細胞が培地から死んでexcludeされると主張しているわけではない。 元論文を押さえれば、BFPが残っていること自体は問題ではありませんね。でも小保方さんはリジェンドで<JAK inhibitor treatment assay eliminated ES cells (Oct4-GFP+) from the culture (a, b).>と書いていて、ES細胞が多能性細胞としての機能を失うという意味に受け止められないことはありませんが、同時に細胞自体が死滅するという意味が残ってしまう。またレフェリーのアドバイス<Presence of ES cells could be excluded by culture in the presence of JAK inhibitor,)も同じです。
[連投5]
この相違は胎膜(fetal membranes)と卵黄嚢(yolk sac)に関する笹井さんの書いた本文と小保方さんの書いたリジェンドにも出現していて、笹井さんは胎膜(fetal membranes)と書いていますね。
>Surprisingly, injected STAP cells contributed not only to the embryo but also to the placenta and fetal membranes (Fig. 1b and Extended Data Fig. 1a–c) in 60% of the chimaeric embryos (Fig. 1c).
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Pub Peerの#12コメント冒頭は以下です。
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#12 Peer 5 commented 5 years ago
commented February 28th, 2014 10:04 AM and accepted February 28th, 2014 10:04 AM
The authors claim that "Surprisingly, injected STAP cells contributed not only to the embryo but also to the placenta and fetal membranes (Fig. 1b and Extended Data Fig. 1a-c) in 60% of the chimaeric embryos (Fig. 1c)." and they show the analysis of chimaeric embryos with placentas at E12.5.
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(続き)I'm sorry to say but that is normal in E12.5 chimaeric embryos produced by injection of wt ES cells into blastocyst, since the placental FETAL blood vessels and yolk sac mesoderm cells are derived from the ES-cell derived epiblast.
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ESのキメラ胎盤と卵黄嚢は光って当たり前だと言ってるわけですね。むしろFigure 1-aのESキメラとされているものの方が光ってないことが不思議だと。
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In that sense what is surprising to me is that in their chimaeras (fig 1a and 1c) produced by injection of GFP expressing ES cells (Rosa26GFP or CAG-GFP) into blastocyst none showed fluorescence in the placenta (fetal blood vessels) or yolk sac (yolk sac mesoderm).
[その意味で、私にとって驚くべきことは、胚盤胞へのGFP発現ES細胞(Rosa26GFPまたはCAG-GFP）のインジェクションによって作られたキメラ（図1aおよび1c）に於いて、胎盤（胎児血管）あるいは卵黄嚢（卵黄嚢中胚葉）がまったく蛍光していないことだ。]
