科研データベース
ttp://kaken.nii.ac.jp/d/p/20062015.ja.html
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卵子による核の初期化機構の解明およびその促進方法の開発
Study of nuclear reprogramming using mammalian oocyte
研究課題番号：20062015
N論のAcknowledgementsでこの予算を使っていることは明示されている。
Financial support for this research was provided by Intramural RIKEN Research Budget (H.O., T.W. and Y.S.), a Scientific Research in Priority Areas (20062015) to T.W.,
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expressed pluripotency-related marker proteins (Oct4, SSEA1, Nanog and E-cadherin; Fig. 2a) and marker genes (Oct4, Nanog, Sox2, Ecat1 (also called Khdc3), Esg1 (Dppa5a), Dax1 (Nrob1) and Rex1 (Zfp42); Fig. 2b and Extended Data Fig. 3a) in a manner comparable to those seen in ES cells. Moderate levels of expression of these pluripotency marker genes were observed on day 3 (Fig. 2b and Extended Data Fig. 3b). Notably, the Oct4-GFP+ cells on day 3, but not on day 7, expressed early haematopoietic marker genes such as Flk1 (also called Kdr) and Tal1 (Extended Data Fig. 3c), indicating that Oct4-GFP+ cells on day 3, as judged by their expression pattern at the population level, were still in a dynamic process of conversion.
これかい？
On day 7, unlike CD45+ cells and like ES cells, low-pH-induced Oct4-GFP+ cells displayed extensive demethylation at the Oct4 and Nanog promoter areas (Fig. 2c), indicating that these cells underwent a substantial reprogramming of epigenetic status in these key genes for pluripotency.
警察が入ってしまえば早いだろうね。
In vitro differentiation assays demonstrated that low-pH-induced Oct4-GFP+ cells gave rise to three-germ-layer derivatives (Fig. 2d) as well as visceral endoderm-like epithelium (Extended Data Fig. 3d). When grafted into mice, low-pH-induced Oct4-GFP+ cell clusters formed teratomas (40%, n = 20) (Fig. 2e and Extended Data Fig. 4a–c) but no teratocarcinomas that persistently contained Oct4-GFP+ cells (n = 50). Because some cellular variation was observed in the signal levels of Oct4-GFP within the clusters, we sorted GFP-strong cells (a major population) and GFP-dim cells (a minor population) by FACS on day 7 and separately injected them into mice. In this case, only GFP-strong cells formed teratomas (Extended Data Fig. 4d).
