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NaïveCult™扩展介质

用于reset naïve人类胚胎干细胞和iPS细胞扩增的细胞培养基
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产品号 #05590_C

用于reset naïve人类胚胎干细胞和iPS细胞扩增的细胞培养基

产品优势

  • 支持reset naïve人类胚胎干细胞和iPS细胞的稳健生长
  • 维持高表达naïve-associated基因的人造血干细胞
  • 兼容转基因不依赖，转基因诱导，胚胎来源重置naïve人类胚胎干细胞和iPS细胞
  • 定义配方包含高质量，预先筛选的成分

产品组分包括

  • NaïveCult™扩展基础培养基，400毫升
  • NaïveCult™5X扩展补充，100毫升
  • NaïveCult™1000X扩展补充，500µL
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Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  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 (Dihydrochloride)

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

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实验数据
产品说明书及文档
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概述

稳健地扩大重置naïve人类胚胎干细胞（ES）和诱导多能干细胞（iPS）细胞使用确定的细胞培养基。

Reset naïve在这种培养基中培养的人类多能干细胞（hPSCs）显示出均匀的圆顶，具有折射边缘的相位明亮的集落，并保持与Reset naïve hPSCs相关的基因表达谱（Guo等）。干细胞报告，2016；郭等。发展,2017;Takashima等人。细胞,2014)。Reset naïve在这种培养基中培养的人造血干细胞能够在重新启动后向所有三种体细胞谱系分化。

基于Austin Smith的t2iL + Gö配方，并根据英国剑桥大学剑桥干细胞研究所的许可开发，该培养基与reset naïve人类胚胎干细胞和iPS细胞兼容NaïveCult™感应套件（目录#05580），转基因诱导的复位人多能干细胞（hPSCs） （Takashima et al. 2014），或胚胎来源的复位人多能干细胞（Guo et al. 2016）。

Subtype
Specialized Media
 
Cell Type
Pluripotent Stem Cells
 
Species
Human
 
Application
Cell Culture, Expansion, Maintenance
 
Brand
NaïveCult
 
Area of Interest
Stem Cell Biology
 

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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.

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
Technical 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

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.

Research Area
Workflow Stages

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Species Human
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