EasySep™小鼠TIL（CD45）正选试剂盒
产品号 #72812_C
PI3K和MEK/ERK通路抑制剂；抑制RasGAP和ERK1
若您需要咨询产品或有任何技术问题，请通过官方电话 400 885 9050 或邮箱 info.cn@stemcell.com 与我们联系
总览
luripotin是一种双靶点抑制剂，可抑制Ras GTP酶激活蛋白（RasGAP, Kd = 98 nM）和胞外信号调节激酶1 (ERK1, Kd = 212 nM)（Chen et al.)。抑制RasGAP，增强了Ras信号中PI3激酶通路的活性，从而促进自我更新，而抑制ERK1则会阻断细胞分化（Chen et al.）。
维持和自我更新
·促进自我更新。在无饲养层细胞、无血清和无白血病抑制因子（LIF）的条件下，维持小鼠胚胎干细胞（ES）处于未分化的多能状态，并可至少传代10代（Chen et al.; Xiong et al.）。
·与LIF一起使用时，可提高从难以建立ES细胞系的小鼠品系中衍生ES细胞的效率(Pieters et al.; Yang et al.)。
细胞类型
多能干细胞
种属
人，小鼠，非人灵长类，其它细胞系，大鼠
应用
扩增，培养
研究领域
干细胞生物学
CAS 编号
839707-37-8
化学式
C₂₇H₂₅F₃N₈O₂
纯度
≥98%
通路
MEK/ERK，PI3K/AKT
靶点
ERK，RasGAP
产品说明书及文档
请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。
应用领域
本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。
相关材料与文献
技术资料 (3)
文献 (4)
Efficient and user-friendly pluripotin-based derivation of mouse embryonic stem cells. Stem cell reviews 2012 SEP
Abstract
Classic derivation of mouse embryonic stem (ES) cells from blastocysts is inefficient, strain-dependent, and requires expert skills. Over recent years, several major improvements have greatly increased the success rate for deriving mouse ES cell lines. The first improvement was the establishment of a user-friendly and reproducible medium-alternating protocol that allows isolation of ES cells from C57BL/6 transgenic mice with efficiencies of up to 75%. A recent report describes the use of this protocol in combination with leukemia inhibitory factor and pluripotin treatment, which made it possible to obtain ES cells from F1 strains with high efficiency. We report modifications of these protocols for user-friendly and reproducible derivation of mouse ES cells with efficiencies of up to 100%. Our protocol involves a long initial incubation of primary outgrowths from blastocysts with pluripotin, which results in the formation of large spherical outgrowths. These outgrowths are morphologically distinct from classical inner cell mass (ICM) outgrowths and can be easily picked and trypsinized. Pluripotin was omitted after the first trypsinization because we found that it blocks attachment of ES cells to the feeder layer and its removal facilitated formation of ES cell colonies. The newly established ES cells exhibited normal karyotypes and generated chimeras. In summary, our user-friendly modified protocol allows formation of large spherical ICM outgrowths in a robust and reliable manner. These outgrowths gave rise to ES cell lines with success rates of up to 100%.
The use of SC1 (Pluripotin) to support mESC self-renewal in the absence of LIF. Journal of visualized experiments : JoVE 2009 JAN
Abstract
Mouse embryonic stem (ES) cells are conventionally cultured with Leukemia Inhibitory Factor (LIF) to maintain self-renewal.(1) However, LIF is expensive and activation of the LIF/JAK/STAT3 pathway is not absolutely required to maintain the self-renewal state.(2) The SC1 small molecule may be an economical alternative to LIF. SC1 functions through dual inhibition of Ras-GAP and ERK1.(3) Illustration of its mechanism of action makes it a useful tool to study the fundamental molecular mechanism of self-renewal. Here we demonstrate the procedure for culturing mouse ES cells in the presence of SC1 and show that they are able to maintain self-renewal in the absence of LIF. Cells cultured with SC1 showed similar morphology compared to cells maintained with LIF. Both exhibited typical mouse ES morphology after five passages. Expression of typical pluripotency markers (Oct4, Sox2, Nanog, and SSEA1) was observed after five passages in the presence of SC1. Furthermore, SC1 caused no overt toxicity on mouse ES cells.
Pluripotin combined with leukemia inhibitory factor greatly promotes the derivation of embryonic stem cell lines from refractory strains. Stem cells (Dayton, Ohio) 2009 FEB
Abstract
Most mouse embryonic stem (ES) cells are derived from a 129 or C57BL/6 background, whereas the derivation efficiency of ES cells is extremely low on certain refractory types of background for which ES cells are highly desired. Here we report an optimized, highly efficient protocol by combining pluripotin, a small molecule, and leukemia inhibitory factor (LIF) for the derivation of mouse ES cells. With this method, we successfully isolated ES cell lines from five strains of mice, with an efficiency of 57% for NOD-scid, 63% for SCID beige, 80% for CD-1, and 100% for two F1 strains from C57BL/6xCD-1. By tracking the Oct4-positive cells in the Oct4-green fluorescent protein embryos in the process of ES cell isolation, we found that pluripotin combined with LIF improved the efficiency of ES cell isolation by selectively maintaining the Oct4-positive cells in the outgrowth. To our knowledge, this is the first report of ES cells being efficiently derived from immunodeficient mice on refractory backgrounds (NOD-scid on a NOD background and SCID beige on a BALB/c background).
更多信息
| 种属 | Human, Mouse, Non-Human Primate, Other, Rat |
|---|---|
| Cas Number | 839707-37-8 |
| Chemical Formula | C₂₇H₂₅F₃N₈O₂ |
| 纯度 | ≥ 98% |
| Target | ERK, RasGAP |
| Pathway | MEK/ERK, PI3K/AKT |
