STEMCELL Technologies NaiveCult NaiveCult Induction Kit
- 研究用
NaïveCult™ Induction Kit (ST-05580) は、プライム状態のヒトES/iPS細胞をリセットし、ナイーブ状態に化学的誘導する導入遺伝子フリーの培地です。
本キットでリセットしたナイーブ型ヒト多能性幹細胞 (hPSC) は、縁が屈折した明るい位相差像の均一なドーム型コロニーを示し、リセットされたナイーブ状態に関連する遺伝子発現プロファイルを維持します (Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014)。
Guo et al. (2017) の組成に基づき、英国ケンブリッジ大学の Cambridge Stem Cell Institute からライセンスを受けて開発された本キットは、mTeSR™1 (ST-85850）または TeSR™-E8™ (ST-05990) で維持した複数のプライム型ヒトES/iPS細胞に対し堅牢に機能します。
キットの使用には、別売のヒストンデアセチラーゼ阻害剤 (HDACi；Sodium Butyrate [ST-72242]または Valproic Acid [ST-72292]) が必要になります。
2018/05/14 12:00の製品情報
本製品は研究目的にのみ使用し、人や動物の医療用・臨床診断用・食品用としては使用しないようにご注意ください。
製品の特長
NaïveCult™ Induction Kitをもちいて、ナイーブ型hPSCをトランスジーンフリーで誘導できます
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).
