アーティクルのTCRの件は以下ね。
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TCR-β chain gene rearrangement analysis
Genomic DNA was extracted from STAP cells and tail tips from chimaeric mice generated with STAP cells derived from CD45+ cells. PCR was performed with 50 ng DNA using the following primers (Dβ2: 5′-GCACCTGTGGGGAAGAAACT-3′ and Jβ2.6: 5′-TGAGAGCTGTCTCCTACTATCGATT-3′) that amplify the regions of the (D)J recombination. The PCR products were subjected to gel electrophoresis in Tris-acetate-EDTA buffer with 1.6% agarose and visualized by staining with ethidium bromide. PCR bands from STAP cells were subjected to sequencing analysis and identified as rearranged genomic fragments of the (D)J recombination.
我々は小保方さんは無実という立場だからね。当然だけどB6-CAG-GFPx129/Svだと
若山さんに言われたからそう書いていて、かつ親の雌雄の違いは見た目では分からないよね。
そして「あるとき」ではなくて2012年の3月に光る胎盤を伴うキメラ胎児を
受け取って、それを若山さんの指示通りに、切片の免染確認でGFPを胎盤に検出しているのが
Letter Extended Data Figure 1 Placental contribution of STAP cellsなんだよな。
*ttps://ipscell.com/2014/02/are-stap-stem-cell-nature-papers-compromised/
Nimorula
February 25, 2014 at 7:19 pm .
In the paper “Bidirectional developmental potential…” besides the duplicated image in figures 1 and 2 there are things really hard to believe about the data they show in figure 1. First they should have done the analysis in chimaeric embryos previous to the chrorio-allantoic fusion (embryos before 6 somites, around E8) to conclude that only STAPs and not ES cells contribute to the placenta. Developing my point: After the allantois (derived from the embryonic mesoderm) fuses with the chorion (extraembryonic) the formation of the labyrinthine placenta occurs. In this, the trophoblast (extraembryonic) associates with the fetal blood vessels (embryonic) that have extensive villous branching. Then in an E12.5 placenta from a chimaera formed from a wildtype blastocyst injected with wildtype reporter Rosa26-GFP or CAG-GFP ES cells you should always find GFP positive cells in the placenta due to the contribution of the fetal blood vessels. Then is not possible that in the control chimaeras in figure 1a they didn’t find Rosa26-GFP cells (as stated in the text) / CAG-GFP (as stated in the figure). The same applies for the fetal membranes, the yolk sac mesoderm is derived from the embryo then in a chimaera you always have ES-derived cells (mesoderm) in the yolk sac. There is no way that control is real. Mild: the long exposure panel of the control embryo was not overexposed since the little piece of embryo in the far right of the panel is not overexposed.
nimorula
February 26, 2014 at 2:21 pm .
p.s. A nice example of chimaeric embryos analysing the contribution of ES cell derived tissue to the placenta in E12.5 embryos can be found in PNAS USA 100(26):15637-15642. (2003) see figure 4.
