NaïveCult™ Induction Kit

Defined medium kit for transgene-free induction of reset naïve human ES and iPS cells

NaïveCult™ Induction Kit

Defined medium kit for transgene-free induction of reset naïve human ES and iPS cells

NaïveCult™ Induction Kit for the generation of reset hPSCs
1 Kit
1,274 USD
Catalog # 05580

Defined medium kit for transgene-free induction of reset naïve human pluripotent stem cells

Product Advantages


  • Transgene-free chemical induction of reset human naïve ES and iPS cells

  • Robust induction across multiple human ES and iPS cell lines

  • Generates hPSCs with high expression of naïve-associated genes

  • Defined formulation containing high-quality, pre-screened components

What's Included

  • NaïveCult™ Induction Basal Medium, 76 mL
  • NaïveCult™ 20X Induction Supplement A, 4 mL
  • NaïveCult™ 1000X Induction Supplement B, 80 µL
  • NaïveCult™ Expansion Basal Medium, 2 x 400 mL
  • NaïveCult™ 5X Induction Supplement C, 200 mL
  • NaïveCult™ 1000X Induction Supplement D, 500 µL
  • NaïveCult™ 1000X Expansion Supplement, 500 µL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

Perform transgene-free chemical induction of reset human naïve embryonic stem (ES) and induced pluripotent stem (iPS) cells from the primed state using a defined cell culture medium.

Reset naïve human pluripotent stem cells (hPSCs) generated using this kit demonstrate uniformly domed, phase-bright colonies with refractive edges and maintain gene expression profiles associated with reset naïve hPSCs (Guo et al. Stem Cell Reports, 2016; Guo et al. Development, 2017; Takashima et al. Cell, 2014).

Based on the formulation by Guo et al. (2017) and developed under license from the Cambridge Stem Cell Institute at the University of Cambridge, UK, this kit works robustly across multiple primed human ES and iPS cells maintained in mTeSR™1 (Catalog #85850) or TeSR™-E8™ (Catalog #05990). Note that NaïveCult™ Induction Kit requires a histone deacetylase inhibitor (HDACi; either Sodium Butyrate (Catalog #72242) or Valproic Acid (Catalog #72292)) that is sold separately.
Subtype
Specialized Media
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Expansion, Maintenance
Brand
NaïveCult
Area of Interest
Stem Cell Biology

Data Figures

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).

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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Technical Manual
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Safety Data Sheet 1
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Safety Data Sheet 7
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Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

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