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
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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
Coating for cultureware
Epigenetic modifier; Inhibits histone deacetylase
Epigenetic modifier; Inhibits histone deacetylase (HDAC)1
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
Compatible antibodies for purity assessment of isolated cells
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
人
应用
细胞培养,扩增,培养
品牌
NaïveCult
研究领域
干细胞生物学
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).
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用于naïve态人胚胎干细胞和iPS细胞扩增的细胞培养基
无饲养层、无动物成分的人胚胎干细胞和诱导多能干细胞维持培养基
细胞分离溶液
cGMP标准、无饲养层的人胚胎干细胞(ES)和诱导多能干细胞(iPS)维持培养基
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