The D3 ES cell line was originally established by Doetschman et al. [12], and its GFP transgenic cell line (129SV(D3)-Tg(ACTB-EGFP)CZ-001-FM260Osb), which was established using pCAG-EGFP [10, 11], was kindly provided by Dr. Masaru Okabe (Osaka University, Osaka, Japan). The E14 ES cell line [7] was derived from the inbred mouse strain 129/Ola in 1985 by Dr. Martin Hooper in Edinburgh (Scotland) and was obtained through Dr. Peter Mombaerts (Rockefeller University). The 129B6F1G1 [13] and BDmt2 [14] are nuclear transfer-derived ES (ntES) cell lines [15] previously established in our laboratory using Sertoli cells of 129B6F1 background with GFP and tail-tip cells of a male BDF1 mouse as donor for nuclear transfer, respectively. Male 129B6F1 mice expressing GFP were generated by mating female 129/Sv mice with male C57BL/6 GFP transgenic mice (double-transgenic mouse line [16] constructed using pCAG-EGFP [10, 11] and Acr3-EGFP [17]).
あれ、デラ、僕の名前で書きこんでるぜ。
問題は４行目意以降だね。
>>
he 129B6F1G1 [13] and BDmt2 [14] are nuclear transfer-derived ES (ntES) cell lines [15] previously established in our laboratory using Sertoli cells of 129B6F1 background with GFP and tail-tip cells of a male BDF1 mouse as donor for nuclear transfer, respectively. Male 129B6F1 mice expressing GFP were generated by mating female 129/Sv mice with male C57BL/6 GFP transgenic mice (double-transgenic mouse line [16] constructed using pCAG-EGFP [10, 11] and Acr3-EGFP [17]).
今はBDmt2は関係ないから、その記述を外せばわかりやすいね。
>>
The 129B6F1G1 [13] ・・・ are nuclear transfer-derived ES (ntES) cell lines [15] previously established in our laboratory
using Sertoli cells of 129B6F1 background with GFP ・・・as donor for nuclear transfer, ・・・.
Male 129B6F1 mice expressing GFP were generated by mating female 129/Sv mice with male C57BL/6 GFP transgenic mice
(double-transgenic mouse line [16] constructed using pCAG-EGFP [10, 11] and Acr3-EGFP [17]).
Jのリジェンドね。FLSの8ラインの４Nキメラのジャームライントランスミッション実験結果ね。
>>
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.
基本はArticle Fifure2のeのテラトーマ写真だね。リジェンドにOct4-GFPと書かれている。
>>
e, Teratoma formation assay of day 7 clusters of Oct4-GFP+ cells. Haematoxylin and eosin staining showed keratinized epidermis (ectoderm), skeletal muscle (mesoderm) and intestinal villi (endoderm), whereas immunostaining showed expression of Tuj1 (neurons), smooth muscle actin and α-fetoprotein. Scale bar, 100 μm.
129B6F1G1 and GR14 are nuclear transfer-
derived ES(ntES) cell lines (Wakayama et al
2001) previously established in our laboratory
using sertoli cell from a 129B6F1 background
With GFP(Ohta ＆ Wakayama 2005) and
tail tip cell of a mail mous
(129BDF2. Wakayama et al 2005) respectively
as donor for nuclear transfer karyotype
analysis revealed that the ES cell lines
Used had the normal karyotype at the
following percentages (number of metaphases
with normal karyotype in parentheses)
46%(12／26)for E14 35%(9／26) for
129B6F1G1 and54%(14／26)for GR14
ここね。
>>
To evaluate whether the iMuSCs could give rise to chimeric mice, a blastocyst injection assay was performed (Fig. 4c). We transferred undifferentiated β-gal+ and GFP-pre-labelled iMuSCs as single cells into BALB/c (Jackson Lab, USA) blastocysts by microinjection following standard procedures[8].
>>
[8].Nagy, A. Manipulating the mouse embryo: a laboratory manual. 3rd edn, (Cold Spring Harbor Laboratory Press, 2003).
ティシュー論文からですわね。
>>
Expression of embryonic-like phenotypes
Expression of the following genes normally associated with the early stage of embryonic development was seen in cells procured from each tissue. The gene profiles that were expressed are recognized as ESC markers and suggestive of pluripotency. Oct4, Nanog, Zfp296, Cripto, Gdf3, Utf1, Ecat1, Esg1, Sox2, and Fgf5 (Figs. 2 and 3) were expressed in cells procured from each tissue source. In contrast, Rex1, Eras, and Cdx2 were not expressed at the mRNA level (Figs. 2 and 3; Supplementary Table S2), suggesting that although the cells analyzed in the study were very similar to ESCs, they were not identical.
レター論文はこちらね。
>>
In quantitative polymerase chain reaction (qPCR) analysis, STAP cells (sorted for strong Oct4-GFP signals) expressed not only pluripotency marker genes but also trophoblast marker genes such as Cdx2 (Fig. 1d and Supplementary Table 1 for primers), unlike ES cells. Therefore, the blastocyst injection result is not easily explained by the idea that STAP cells are composed of a simple mixture of pluripotent cells (Oct4+Cdx2−) and trophoblast-stem-like cells (Oct4−Cdx2+).
