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STEMdiff™星形细胞成熟试剂盒

从人胚胎干细胞和iPS细胞衍生的星形胶质细胞前体生成皮质型星形胶质细胞的成熟试剂盒
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产品号 #100-0016_C

从人胚胎干细胞和iPS细胞衍生的星形胶质细胞前体生成皮质型星形胶质细胞的成熟试剂盒

产品优势

  • 支持高效生成功能性星形胶质细胞
  • STEMdiff™SMADi神经诱导试剂盒优化神经祖细胞分化
  • 使来自多种人类胚胎干细胞和iPS细胞系的皮质型星形细胞前体可再生成熟

产品组分包括

  • STEMdiff™星形细胞成熟基础培养基，80 mL
  • STEMdiff™星形细胞成熟补充剂A， 20ml
  • STEMdiff™星形细胞成熟补充剂B， 1ml
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Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. STEMdiff™ Neural Rosette Selection Reagent
    STEMdiff™ Neural Rosette Selection Reagent

    Enzyme-free reagent for the selective detachment of neural rosettes

  2. STEMdiff™ SMADi Neural Induction Kit
    STEMdiff™ SMADi Neural Induction Kit

    Serum-free medium kit for highly efficient SMAD inhibition-mediated neural induction of human ES and iPS cells

  3. STEMdiff™ Astrocyte Differentiation Kit
    STEMdiff™ Astrocyte Differentiation Kit

    Serum-free differentiation kit for generating astrocyte precursors from hPSC-derived neural progenitor cells

  4. Human iPSC-Derived Neural Progenitor Cells
    Human iPSC-Derived Neural Progenitor Cells

    Frozen human neural progenitor cells differentiated from the human induced pluripotent stem cell line, SCTi003-A

  5. 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

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实验数据
产品说明书及文档
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概述

STEMdiff™星形胶质细胞成熟试剂盒用于使用STEMdiff™星形胶质细胞分化试剂盒（目录#100-0016），从人类多能干细胞（hPSCs）衍生的星形胶质细胞前体快速有效地生成皮质型星形胶质细胞。使用该系统，高纯度的星形胶质细胞群(平均> 70%为s100b阳性，> 60%为gmap阳性；< 15%的双皮质素阳性神经元)可以在短短7周内从hPSCs中生成，并且可以在培养中长期维持。使用这些产品衍生的细胞是模拟人类神经发育和疾病、药物筛选、毒性测试和细胞治疗验证的通用工具。

Subtype
Specialized Media
 
Cell Type
Astrocytes, Neural Cells, PSC-Derived
 
Species
Human
 
Application
Cell Culture, Differentiation, Functional Assay
 
Brand
STEMdiff
 
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Neuroscience
 

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Data Figures

Experimental Protocol Schematic for STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits (Embryoid Body Protocol)

Figure 1. Schematic for the Embryoid Body Protocol

Cortical-type astrocytes can be generated from astrocyte precursors after 20 days in STEMdiff™ Astrocyte Differentiation Medium. For differentiation of precursors from embryonic and induced pluripotent stem cells, see the PIS.

Experimental Protocol Schematic for STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits (Monolayer Protocol)

Figure 2. Schematic for the Monolayer Protocol

Cortical-type astrocytes can be generated from astrocyte precursors after 21 days in STEMdiff™ Astrocyte Differentiation Medium. For differentiation of precursors from embryonic and induced pluripotent stem cells, see the PIS.

Culturing PSCs in STEMdiff™ SMADi Neural Induction Kit and STEMdiff™ Astrocyte Differentiation and Maturation Kits Yields Cortical-Type Astrocytes

Figure 3. Cortical-Type Astrocytes Are Generated After Culture in STEMdiff™ Astrocyte Differentiation and Maturation Kits

NPCs generated from hPSCs in TeSR™-E8™ using the STEMdiff™ SMADi Neural Induction Kit embryoid body (EB) protocol were differentiated and matured to cortical-type astrocytes using the STEMdiff™ Astrocyte Differentiation and Maturation Kits. Cortical-type astrocytes were formed after iPS cell-derived NPCs were cultured with the STEMdiff™ Astrocyte Differentiation Kit for 3 weeks and STEMdiff™ Astrocyte Maturation Kit for 3 weeks. (A) Nuclei are labeled with DAPI (gray). The resulting cultures contain a highly pure population of astrocytes, which are (B) more than 60% GFAP-positive (green) and (C) more than 70% S100B-positive (magenta), with (D) fewer than 15% neurons (DCX-positive cells, cyan). Scale bar = 100 μm.

Figure 4. STEMdiff™ Astrocyte Kits Generate Cells Expressing Expected Levels of Genes Characteristic for Astrocytes

Embryonic stem and induced pluripotent stem cells from a variety of lines (n = 6, maintained in mTeSR™1 or TeSR™-E8™) were differentiated to NPCs using the STEMdiff™ SMADi Neural Induction Kit embryoid body protocol. Cells were then grown in STEMdiff™ Astrocyte Differentiation Kit for 3 weeks followed by STEMdiff™ Astrocyte Maturation Kit for 3 weeks prior to analysis. Expression levels were measured by quantitative PCR (qPCR) and normalized to hPSC controls relative to housekeeping genes 18S and TBP.

Figure 5. PSC-Derived Astrocytes and Neurons Can Be Co-Cultured to Model Cell-Cell Interactions In Vitro

NPCs generated from the H1 cell line were differentiated to astrocytes using STEMdiff™ Astrocyte Differentiation and Maturation Kits. H9 cell-derived NPCs were differentiated to forebrain-type neurons using STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits. For co-culture, matured astrocytes were seeded onto forebrain neurons that had been in STEMdiff™ Forebrain Neuron Maturation Medium for at least one week. Co-cultures were then switched to STEMdiff™ Forebrain Neuron Maturation Medium the following day and for the remaining co-culture. (A) Neurons cultured alone, following the co-culture feeding schedule, are labeled with DCX (green). (B) DCX-positive neurons (green) and astrocytes (GFAP, red) can be co-cultured for at least 1 - 2 weeks prior to analysis. For a detailed co-culture protocol, please see the Methods Library.

Figure 6. PSC-Derived Neurons Survive and Mature when Co-Cultured with PSC-Derived Astrocytes

NPCs generated from the STiPS-R038 cell line were differentiated to astrocytes using STEMdiff™ Astrocyte Differentiation and Maturation Kits. STiPS-M001 cell-derived NPCs were differentiated to forebrain-type neurons using STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits. After co-culture for one week, neurons (A) had significantly increased neurite outgrowth as measured on MAP2-positive neurons with the NeuriteTracer plugin for ImageJ (M Pool et al. J Neurosci Methods, 2008) and (B) were more numerous than neurons cultured alone using the same feeding schedule. *, p < 0.05; **, p < 0.01.

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
STEMdiff™ Astrocyte Maturation Kit
Catalog #
100-0016
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
STEMdiff™ Astrocyte Maturation Kit
Catalog #
100-0016
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
STEMdiff™ Astrocyte Maturation Kit
Catalog #
100-0016
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
STEMdiff™ Astrocyte Maturation Kit
Catalog #
100-0016
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
STEMdiff™ Astrocyte Maturation Kit
Catalog #
100-0016
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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