彼女はヴァカンティ研究室で逆転写PCR実験によってOct4蛋白を発見してから、
ヴァカンティに認められ、その後ラボの全員の協力を得てあのティシュー誌に
載せられている各マーカー蛋白のゲル上の電気泳動バンドの図表を作ったんだね。
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Although the spheres in this study described were capable of generating teratoma-like tissue, the transplanted cells did not form large teratomas as did ES cells, nor they did express Eras in vitro as ESCs,31 which suggests that the teratoma-like tissue they generate may be very different from true teratomas generated from ESCs. In addition, the cells studied did not express the trophectoderm marker Cdx232 or also associated with ESCs. These differences of gene expression pattern may explain the differences of the biological function between ESCs and adult stem cells in this study.
レター論文の参照は以下ね。
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16.
Yang, J. et al. Stat3 activation is limiting for reprogramming to ground state pluripotency. Cell Stem Cell 7, 319–328 (2010)
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ど素人には手が回らないね。あはははは。
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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).
レター論文のa JAK inhibitor <16>の参照がヤンさんの論文だけど今ざっと
目を通した限りでJAK阻害剤処置でESが取り除かれるとは書かれてないように見える。
にも拘わらず、Extended Data Fig. 5にはその実証証明がある。ヤンさんの論文は
今見つけたばかりだからもうちょっと詳しく調べないと分からないね。
ヤンさんの論文の小保方さんが言ってる部分は以下だわね。
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Results
Lif Increases the Efficiency of EpiSC Reprogramming
We compared the frequency of Epi-iPSC generation in the presence or absence of Lif. We used embryo-derived Oct4-GFP (O4G) reporter EpiSCs stably transfected with expression constructs for Klf4 or Nanog (Guo et al., 2009). On transfer from activin plus Fgf into 2i with Lif, these cells produced GFP-positive iPSC colonies at a frequency of 0.5%–1% (Guo et al., 2009, Silva et al., 2009). Without Lif or in the presence of a Jak inhibitor, this yield was reduced several fold (Figure 1A ). To test whether the effect of Lif is due simply to increased efficiency of iPSC self-renewal, we plated reprogrammed Epi-iPSCs in 2i with or without Lif. We observed no significant difference in numbers of colonies formed or their undifferentiated phenotype (Figure 1B).
<Without Lif or in the presence of a Jak inhibitor, this yield was
reduced several fold (Figure 1A ). >のところね。図の1-Aは棒グラフだけど
小保方さんがLetter Extendes Data5で示した写真と同じ結果ね。
