NaïveCult™ Reset Naïve Human ES/iPS Cell Expansion Medium

总览

使用成分明确的细胞培养基，可高效扩增naïve态人类胚胎干细胞（ES）和诱导多能干细胞（iPS）。在该培养基中培养naïve态人多能干细胞（hPSCs）具有折射边缘的均匀圆顶、相位明亮的集落，并保持与重置Naive hPSC相当的基因表达谱（参考文献：Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014）。

在重置为始发态（re-priming）后，这些细胞可实现三胚层方向的分化，具备向所有三种体细胞谱系分化的能力。

该培养基基于Austin Smith的t2iL + Gö配方，并获得英国剑桥大学剑桥干细胞研究所的授权，其兼容广泛，适用于使用NaïveCult™诱导试剂盒（产品号：#05580）获得的重置naïve人ES/iPS细胞、转基因诱导的重置型hPSC（Takashima 等，2014）以及胚胎来源的重置型hPSC（Guo 等，2016）

在重置为始发态（re-priming）后，这些细胞可实现三胚层方向的分化，具备向所有三种体细胞谱系分化的能力。

该培养基基于Austin Smith的t2iL + Gö配方，并获得英国剑桥大学剑桥干细胞研究所的授权，其兼容广泛，适用于使用NaïveCult™诱导试剂盒（产品号：#05580）获得的重置naïve人ES/iPS细胞、转基因诱导的重置型hPSC（Takashima 等，2014）以及胚胎来源的重置型hPSC（Guo 等，2016）

在重置为始发态（re-priming）后，这些细胞可实现三胚层方向的分化，具备向所有三种体细胞谱系分化的能力。

该培养基基于Austin Smith的t2iL + Gö配方，并获得英国剑桥大学剑桥干细胞研究所的授权，其兼容广泛，适用于使用NaïveCult™诱导试剂盒（产品号：#05580）获得的重置naïve人ES/iPS细胞、转基因诱导的重置型hPSC（Takashima 等，2014）以及胚胎来源的重置型hPSC（Guo 等，2016）

在重置为始发态（re-priming）后，这些细胞可实现三胚层方向的分化，具备向所有三种体细胞谱系分化的能力。

该培养基基于Austin Smith的t2iL + Gö配方，并获得英国剑桥大学剑桥干细胞研究所的授权，其兼容广泛，适用于使用NaïveCult™诱导试剂盒（产品号：#05580）获得的重置naïve人ES/iPS细胞、转基因诱导的重置型hPSC（Takashima 等，2014）以及胚胎来源的重置型hPSC（Guo 等，2016）

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

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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).