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NaïveCult™感应套件

定义培养基试剂盒，用于无转基因诱导重置naïve人ES和iPS细胞
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产品号 #05580_C

定义培养基试剂盒，用于无转基因诱导重置naïve人ES和iPS细胞

产品优势

  • 重置人类naïve ES和iPS细胞的无转基因化学诱导
  • 对多种人类胚胎干细胞和iPS细胞系的强诱导
  • 生成高表达naïve-associated基因的人造血干细胞
  • 定义配方包含高质量，预先筛选的成分

产品组分包括

  • NaïveCult™诱导基础培养基，76 mL
  • NaïveCult™20X诱导补充A， 4ml
  • NaïveCult™1000X诱导补充B， 80µL
  • NaïveCult™扩展基础培养基，2 x 400 mL
  • NaïveCult™5X诱导补充C， 200毫升
  • NaïveCult™1000X感应补充D， 500µL
  • 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. Sodium Butyrate
    Sodium Butyrate

    Epigenetic modifier; Inhibits histone deacetylase

  3. Valproic Acid (Sodium Salt)
    Valproic Acid (Sodium Salt)

    Epigenetic modifier; Inhibits histone deacetylase (HDAC)1

  4. Y-27632 (Dihydrochloride)
    Y-27632 (Dihydrochloride)

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

总览
实验数据
产品说明书及文档
应用领域
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概述

使用确定的细胞培养基对重置的人类naïve胚胎干细胞（ES）和诱导多能干细胞（iPS）细胞进行无转基因化学诱导。

Reset naïve使用该试剂盒生成的人类多能干细胞（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)。

基于Guo等人（2017）的配方，并根据英国剑桥大学剑桥干细胞研究所的许可开发，该试剂盒可在保存的多个引物人类胚胎干细胞和iPS细胞中稳定工作mTeSR™1（目录#85850）或TeSR™e8™(目录# 05990)。注意NaïveCult™诱导试剂盒需要组蛋白去乙酰化酶抑制剂(HDACi；要么丁酸钠（目录#72242）或丙戊酸（目录#72292）)，这是单独出售。

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™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Technical Manual
Product Name
NaïveCult™ Induction Kit
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
Lot #
All
Language
English
Document Type
Safety Data Sheet 7
Product Name
NaïveCult™ Induction Kit for the generation of reset hPSCs
Catalog #
05580
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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