ま、そういうことで。
Here we report a unique cellular reprogramming phenomenon, called stimulus-triggered acquisition of pluripotency (STAP), which requires neither nuclear transfer nor the introduction of transcription factors. In STAP, strong external stimuli such as a transient low-pH stressor reprogrammed mammalian somatic cells, resulting in the generation of pluripotent cells. Through real-time imaging of STAP cells derived from purified lymphocytes, as well as gene rearrangement アナリシス, we found that committed somatic cells give rise to STAP cells by reprogramming rather than selection. STAP cells showed a substantial decrease in DNA methylation in the regulatory regions of pluripotency marker genes. Blastocyst injection showed that STAP cells efficiently contribute to chimaeric embryos and to offspring via germline transmission. We also demonstrate the derivation of robustly expandable pluripotent cell lines from STAP cells. Thus, our findings indicate that epigenetic fate determination of mammalian cells can be markedly converted in a context-dependent manner by strong environmental cues.
1. real-time imaging of STAP cells derived from purified lymphocytes
2.gene rearrangement アナリシス,
3.reprogramming rather than selection
4.DNA methylation in the regulatory regions of pluripotency marker genes
5.Blastocyst injection showed that STAP cells efficiently contribute to chimaeric embryos
6.to offspring via germline transmission
7.We also demonstrate the derivation of robustly expandable pluripotent cell lines from STAP cells.
8.Thus, our findings indicate that epigenetic fate determination of mammalian cells can be markedly converted in a context-dependent manner by strong environmental cues.
この出だしって誰が書いたのかな。笹井さんなのかな。
In the キャナライゼイション view of Waddington’s epigenetic landscape, fates of somatic cells are progressively determined as cellular differentiation proceeds, like going downhill. It is generally believed that reversal of differentiated status requires artificial physical or genetic manipulation of nuclear function such as nuclear transfer, or the introduction of multiple transcription factors. Here we investigated the question of whether somatic cells can undergo nuclear reprogramming simply in response to external triggers without direct nuclear manipulation. This type of situation is known to occur in plants—drastic environmental changes can convert mature somatic cells (for example, dissociated carrot cells) into immature blastema cells, from which a whole plant structure, including stalks and roots, develops in the presence of auxins.
ま、そうかな。
A challenging question is whether animal somatic cells have a similar potential that emerges under special conditions. Over the past decade, the presence of pluripotent cells (or closely relevant cell types) in adult tissues has been a matter of debate, for which conflicting conclusions have been reported by various groups. However, no study so far has proven that such pluripotent cells can arise from differentiated somatic cells.
あっ、上私ね。ドンドンいこう。
CD45+ cells were sorted by fluorescence-activated cell sorting (FACS) from the lymphocyte fraction of postnatal spleens (1-week old) of C57BL/6 mice carrying an Oct4-gfp transgene, and were exposed to various types of strong, transient, physical and chemical stimuli (described below). We examined these cells for activation of the Oct4 promoter after culture for several days in suspension using DMEM/F12 medium supplemented with leukaemia inhibitory factor (LIF) and B27 (hereafter called LIF+B27 medium).
この培養中以降にコンタミさせた者が居る。本人で無い限り。
Among the various perturbations, we were particularly interested in low-pH perturbations for two reasons. First, as shown below, low-pH treatment turned out to be most effective for the induction of Oct4. Second, classical experimental embryology has shown that a transient low-pH treatment under ‘sublethal’ conditions can
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alter the differentiation status of tissues. Spontaneous neural conversion from salamander animal caps by soaking the tissues in citrate-based acidic medium below pH 6.0 has been demonstrated previously.
Without exposure to the stimuli, none of the cells sorted with CD45 expressed Oct4-GFP regardless of the culture period in LIF+B27 medium. In contrast, a 30-min treatment with low-pH medium (25-min incubation followed by 5-min centrifugation; Fig. 1a; the most effective range was pH 5.4–5.8; Extended Data Fig. 1a) caused the emergence of substantial numbers of spherical clusters that expressed Oct4-GFP in day-7 culture (Fig. 1b). Substantial numbers of GFP+ cells appeared in all cases performed with neonatal splenic cells (n = 30 experiments).
ＥＳのコンタミとは別問題であり得るんだよな。ここが困る。
＜Substantial numbers of GFP+ cells appeared in all cases performed
with neonatal splenic cells (n = 30 experiments).＞ってとこほとんど
自家蛍光だったと相沢さんが報告している。
