NaïveCult™ Expansion Medium

Defined cell culture medium for expansion of reset naïve human ES and iPS cells
NaïveCult™ Expansion Medium

Defined cell culture medium for expansion of reset naïve human embryonic stem (ES) and induced pluripotent stem (iPS) cells.

1 Kit
Catalog # 05590
389 USD
Required Products
  1. 0.1% Gelatin in Water
    0.1% Gelatin in Water

    Coating for cultureware

  2. DMEM/F-12 with 15 mM HEPES
    DMEM/F-12 with 15 mM HEPES

    Dulbecco's Modified Eagle's Medium/Nutrient Ham's Mixture F-12 (DMEM/F-12) with 15 mM HEPES buffer

  3. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

Overview

NaïveCult™ Expansion Medium is a defined medium for the robust expansion of reset naïve human embryonic stem (ES) and induced pluripotent stem (iPS) cells. This medium was developed under license from the Cambridge Stem Cell Institute at the University of Cambridge, UK based on the t2iL + Gö formulation developed by Austin Smith. It is compatible with reset naïve human ES and iPS cells derived using NaïveCult™ Induction Kit (Catalog #05580), transgene-induced reset human pluripotent stem cells (hPSCs) (Takashima et al.), or embryo-derived reset hPSCs (Guo et al. 2016). Reset naïve human hPSCs cultured in NaïveCult™ Expansion Medium demonstrate uniformly domed, phase-bright colonies with refractive edges and maintain gene expression profiles associated with reset naïve hPSCs (Guo et al. 2016; Guo et al. 2017; Takashima et al.).


References
Takashima Y et al. (2014) Cell 158(6): 1254–69.
Guo G et al. (2016) Stem Cell Reports 6(4): 437–46.
Guo G et al. (2017) Development 144(15): 2748–63.
Advantages
• Supports robust growth of reset naïve human ES and iPS cells
• Maintains hPSCs with high expression of naïve-associated genes
• Compatible with transgene-independent, transgene-induced, and embryo-derived reset naïve human ES and iPS cells
• Defined formulation containing high-quality, pre-screened components
Components
  • NaïveCult™ Expansion Basal Medium, 400 mL
  • NaïveCult™ 5X Expansion Supplement, 100 mL
  • NaïveCult™ 1000X Expansion Supplement, 500 µL
Subtype
Specialized Media
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Expansion, Maintenance
Brand
NaïveCult
Area of Interest
Stem Cell Biology

Related Products

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
NaïveCult™ Expansion Medium
Catalog #
05590
Lot #
All
Language
English
Document Type
Manual
Product Name
NaïveCult™ Expansion Medium
Catalog #
05590
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
NaïveCult™ Expansion Medium
Catalog #
05590
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
NaïveCult™ Expansion Medium
Catalog #
05590
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
NaïveCult™ Expansion Medium
Catalog #
05590
Lot #
All
Language
English

Educational Materials(4)

Brochure
NaïveCult™ Media System
Brochure
Products for Human Pluripotent Stem Cells
Wallchart
Stem Cell States: Naive to Primed Pluripotency
Webinar
24:11
Human Naïve-Like Pluripotent Stem Cell Generation and Maintenance

Product 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.

Data and Publications

Data

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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