相沢さんが報告しているのは環境の劣悪さ、時間の無さ、スフィア塊はできる。
GFP蛍光は弱い。キメラはできないというものだね。テラトーマに関しては以下だ。
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Previous studies also examined the pluripotency of purported STAP cells by their potency to generate teratomas in immune-deficient mice. However, more than 105 cells are required to form teratoma subcutaneously in the flank of an immune-deficient mouse using ES or EC (embryo carcinoma) cells, and the process takes about one month. No teratoma formation was examined in the present study, since the frequency of green fluorescent cell aggregates was low and time was limited. Teratoma formation under the kidney capsule, which also takes about two months using blastocyst embryos, was also not examined.
丹羽さんはイントロでさりげなく書いてるんだよね。無論多義的な読み方をすればだけどね。
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Full reprograming of somatic cells results in the acquisition of the ability to give rise to an entire organism, or totipotency; this can be achieved by somatic cell nuclear transfer.
丹羽さんの報告書のマテメソにある以下は相沢さんの研究だわよね。
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R26R-H2B-EGFP transgenic mouse (Rosa-GFP) line was generated by Laboratory for Animal Resources and Genetic Engineering (LARGE), RIKEN CDB17.
ここもとてもおかしい場所でね。彼らは<Article のメソッド>だけを言ってるんだが
レターにもこのマウスが出ているよね。例のFigure1のaなんだぜ。ES細胞だ。
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Here we have investigated the unique nature of STAP cells, focusing on their differentiation potential into the two major categories (embryonic and placental lineages) of cells in the blastocyst. We became particularly interested in this question after a blastocyst injection assay revealed an unexpected finding. In general, progeny of injected ES cells are found in the embryonic portion of the chimaera, but rarely in the placental portion (Fig. 1a; shown with Rosa26-GFP). 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).
アーティクルの部分はこれね。
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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.
