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NaïveCult™诱导试剂盒

无转基因诱导人ES细胞和iPS细胞重置为原始状态的成分明确型培养基试剂盒

产品号 #(选择产品)

产品号 #05580_C

无转基因诱导人ES细胞和iPS细胞重置为原始状态的成分明确型培养基试剂盒

产品优势

  • 采用无转基因的化学方法诱导人原始态ES/iPS细胞重置
  • 对多种人ES和iPS细胞系均具有稳定、可靠的诱导效果
  • 可获得高表达原始态相关基因的人多能干细胞(hPSCs)
  • 成分明确的培养体系,含有经过严格预筛选的高质量试剂成分

产品组分包括

  • NaïveCult™诱导基础培养基,76 mL;
  • NaïveCult™20X诱导添加物 A, 4 mL;
  • NaïveCult™1000X诱导添加物 B, 80 µL;
  • NaïveCult™扩增基础培养基,2 x 400 mL;
  • NaïveCult™5X诱导添加物 C, 200 mL;
  • NaïveCult™1000X扩增添加物 D, 500 µL;
  • NaïveCult™1000X扩展添加物,500 µL
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总览

使用本成分明确的细胞培养基,可对处于primed状态的人胚胎干细胞(ES)和诱导多能干细胞(iPS)进行无转基因的化学诱导重置,将其高效转化为原始态人多能干细胞(naïve hPSCs)。

通过本试剂盒获得的重置原始态hPSCs表现出均一的圆顶状、相位明亮、边缘清晰的克隆形态,并维持与重置原始态hPSCs一致的基因表达谱(参考文献:Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014)。

本试剂盒基于 Guo et al.(2017)发表的配方开发,并由英国剑桥大学干细胞研究所授权,适用于多种使用mTeSR™1培养基(产品号 #85850)或TeSR™-E8™培养基(产品号 #05990)维持的primed人ES和iPS细胞,表现出良好的通用性和稳定性。

请注意:NaïveCult™诱导试剂盒使用过程中需联合使用组蛋白去乙酰化酶抑制剂(HDACi),如Sodium Butyrate(产品号 #72242)或Valproic Acid(产品号 #72292),需单独购买。

通过本试剂盒获得的重置原始态hPSCs表现出均一的圆顶状、相位明亮、边缘清晰的克隆形态,并维持与重置原始态hPSCs一致的基因表达谱(参考文献:Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014)。

本试剂盒基于 Guo et al.(2017)发表的配方开发,并由英国剑桥大学干细胞研究所授权,适用于多种使用mTeSR™1培养基(产品号 #85850)或TeSR™-E8™培养基(产品号 #05990)维持的primed人ES和iPS细胞,表现出良好的通用性和稳定性。

请注意:NaïveCult™诱导试剂盒使用过程中需联合使用组蛋白去乙酰化酶抑制剂(HDACi),如Sodium Butyrate(产品号 #72242)或Valproic Acid(产品号 #72292),需单独购买。

通过本试剂盒获得的重置原始态hPSCs表现出均一的圆顶状、相位明亮、边缘清晰的克隆形态,并维持与重置原始态hPSCs一致的基因表达谱(参考文献:Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014)。

本试剂盒基于 Guo et al.(2017)发表的配方开发,并由英国剑桥大学干细胞研究所授权,适用于多种使用mTeSR™1培养基(产品号 #85850)或TeSR™-E8™培养基(产品号 #05990)维持的primed人ES和iPS细胞,表现出良好的通用性和稳定性。

请注意:NaïveCult™诱导试剂盒使用过程中需联合使用组蛋白去乙酰化酶抑制剂(HDACi),如Sodium Butyrate(产品号 #72242)或Valproic Acid(产品号 #72292),需单独购买。

通过本试剂盒获得的重置原始态hPSCs表现出均一的圆顶状、相位明亮、边缘清晰的克隆形态,并维持与重置原始态hPSCs一致的基因表达谱(参考文献:Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014)。

本试剂盒基于 Guo et al.(2017)发表的配方开发,并由英国剑桥大学干细胞研究所授权,适用于多种使用mTeSR™1培养基(产品号 #85850)或TeSR™-E8™培养基(产品号 #05990)维持的primed人ES和iPS细胞,表现出良好的通用性和稳定性。

请注意:NaïveCult™诱导试剂盒使用过程中需联合使用组蛋白去乙酰化酶抑制剂(HDACi),如Sodium Butyrate(产品号 #72242)或Valproic Acid(产品号 #72292),需单独购买。

亚型
专用培养基
 
细胞类型
多能干细胞
 
种属

 
应用
细胞培养,扩增,培养
 
品牌
NaïveCult
 
研究领域
干细胞生物学
 

实验数据

Figure 1. Schematic of Reversion of Primed to Naïve hPSCs

Primed hPSCs, grown either in mTeSR™1 or TeSR™-E8™ on Corning® Matrigel®, are plated as single cells on inactivated murine embryonic fibroblasts (iMEFs) and treated with Rho-kinase inhibition (10 μM Y-27632) for 24 hours in hypoxic conditions. On day 1, medium is changed to Induction Medium 1 supplemented with a histone deacetylase inhibitor (HDACi) and cells are cultured for 3 days with daily medium changes. On day 4, medium is changed to Induction Medium 2 and cells are cultured until day 11 with daily medium changes. On day 11, hPSCs are passaged as single cells onto fresh iMEFs and subsequently passaged every 3 - 4 days until the end of passage 2 in Induction Medium 2. From passage 3, hPSCs are cultured in Induction Medium 3. Background differentiation will decrease between passage 3 and 8. At this time cells can be transferred into NaïveCult™ Expansion Medium for long-term maintenance and expansion.

Figure 2. Human ES and iPS Cells Can Be Reverted to a Naïve State

Representative images of human (A) passage-7 H9 ES cells and (B) passage-9 WLS-1C iPS cells that were reverted to a naïve state using the NaïveCult™ Induction Kit and subsequently cultured in NaïveCult™ Expansion Medium. During reversion, colonies change from a flat morphology to a tightly packed, domed morphology with refractive edges characteristic of naïve-state hPSCs.

Figure 3. hPSCs Cultured in the NaïveCult™ Media System Express High Levels of Factors Associated with Naïve hPSCs

Gene expression profile of human (A) H9 ES cells and (B) WLS-1C iPS cells reverted using the NaïveCult™ Induction Kit and maintained in NaïveCult™ Expansion Medium.

Figure 4. Reset Naïve Human PSCs Cultured in NaïveCult™ Expansion Medium are Capable of Tri-Lineage Differentiation Following Re-Priming

Reset naïve human iPS cells cultured in NaïveCult™ Expansion Medium were re-primed in mTeSR™1 and differentiated to all three somatic lineages. (A) Representative flow cytometry plot of human WLS-1C iPS cells differentiated using the STEMdiff™ Definitive Endoderm Kit (Catalog #05110) demonstrating >90% CXCR4+/SOX17+ definitive endodermal progenitors. (B) Representative flow cytometry plot of human WLS-1C iPS cells differentiated using STEMdiff™ Mesodermal Induction Medium (Catalog #05220) demonstrating >80% Brachyury+/OCT4- mesodermal progenitors. (C) Representative immunofluorescence image of PAX6 (green) and SOX1 (red) double positive neural progenitors derived from human WLS-1C iPS cells differentiated using the STEMdiff™ SMADi Neural Induction Kit (Catalog #08581).

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。

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种属 Human
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