STEMdiff™ Forebrain Neuron Differentiation Kit

Differentiation kit for the generation of neuronal precursors from human ES and iPS cell-derived neural progenitor cells
STEMdiff™ Forebrain Neuron Differentiation Kit

Differentiation kit for the generation of neuronal precursors from human ES and iPS cell-derived neural progenitor cells

1 Kit
Catalog # 08600
268 USD
Required Products
  1. STEMdiff™ Neural Rosette Selection Reagent
    STEMdiff™ Neural Rosette Selection Reagent

    Enzyme-free reagent for the selective detachment of neural rosettes

  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. 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

  4. STEMdiff™ Forebrain Neuron Maturation Kit
    STEMdiff™ Forebrain Neuron Maturation Kit

    Maturation kit for generation of functional neurons from human ES and iPS cell-derived neuronal precursor cells

Overview

The STEMdiff™ Forebrain Neuron Differentiation Kit is used in conjunction with the STEMdiff™ Forebrain Neuron Maturation Kit (Catalog #08605) to generate a mixed population of forebrain-type (FOXG1-positive) neurons from neural progenitor cells derived from human pluripotent stem cells. This kit is optimized to work with STEMdiff™ SMADi Neural Induction Kit, which supplies the appropriate neural progenitor cells. Neurons derived using these products are versatile tools for modeling human neurological development and disease, drug screening, toxicity testing, and cell therapy validation.
Advantages
• Defined and serum-free
• Supports highly efficient generation of functional neurons from hPSC-derived neuronal precursors
• Produces a highly pure population (≥ 90% neurons) of mixed excitatory and inhibitory neurons that can be maintained long-term in culture
• Optimized for differentiation of neuronal progenitor cells generated using STEMdiff™ SMADi Neural Induction Kit
• Supports neuronal activity for physiologically relevant results
• Enables reproducible maturation of neuronal precursors derived from multiple human ES and iPS cell lines
Components
  • STEMdiff™ Forebrain Neuron Differentiation Basal Medium, 80 mL
  • STEMdiff™ Forebrain Neuron Differentiation Supplement, 20 mL
Subtype
Specialized Media
Cell Type
Neural Cells, PSC-Derived, Neural Stem and Progenitor Cells
Species
Human
Application
Cell Culture, Differentiation
Brand
STEMdiff
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Neuroscience
Formulation
Serum-Free, Chemically Defined

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
STEMdiff™ Forebrain Neuron Differentiation Kit
Catalog #
08600
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
STEMdiff™ Forebrain Neuron Differentiation Kit
Catalog #
08600
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
STEMdiff™ Forebrain Neuron Differentiation Kit
Catalog #
08600
Lot #
All
Language
English

Educational Materials(14)

Brochure
Products for Human Pluripotent Stem Cells
Brochure
2019-2020 Cell Culture Training Catalog
Brochure
STEMCELL Neural Product Portfolio
Brochure
hPSC-Derived Neural Cell Research
Special Protocol
How to Co-Culture Human Pluripotent Stem Cell (hPSC)-Derived Forebrain Neurons and Astrocytes
Special Protocol
How to Co-Culture Human Pluripotent Stem Cell (hPSC)-Derived Forebrain Neurons and Microglia
Wallchart
Cell-Reprogramming Technology and Neuroscience
Wallchart
Derivation and Applications of Human Pluripotent Stem Cells
Wallchart
Directed Differentiation of Pluripotent Stem Cells
Webinar
47:18
Madeline Lancaster on Brain Organoids: Modeling Human Brain Development in a Dish
Webinar
42:10
Development, Compatibility, and Applications of mTeSR™ Plus; an Enhanced Medium for the Maintenance of Human Pluripotent Stem Cells (hPSCs)
Webinar
31:35
Neuronal Phenotyping of an iPS Cell Model of Rett Syndrome
Webinar
23:25
Modeling Neurodegenerative Diseases with STEMdiff™ Pluripotent Stem Cell-Derived Neural Culture Systems
Webinar
25:30
Brains in a Dish: Using Cerebral Organoids to Study Human Brain Development and Disease
Load More Educational Materials

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

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

Figure 1. Schematic for the Embryoid Body Protocol

Forebrain-type neural precursors can be generated in 18 - 19 days from hPSC-derived neural progenitor cells (NPCs) after selecting neural rosettes from replated embryoid bodies. For the maturation of precursors to forebrain-type neurons, see the PIS.

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

Figure 2. Schematic for the Monolayer Protocol

Forebrain-type neural precursors can be generated from neural progenitor cell (NPC) monolayers derived from embryonic and induced pluripotent stem cells after three single-cell passages. For the maturation of precursors to forebrain-type neurons, see the PIS.

Forebrain-Type Neurons Are Generated from Neural Progenitor Cells After Culture in STEMdiff™ SMADi Neural Induction Kit and STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits

Figure 3. Forebrain-Type Neurons Are Generated After Culture in STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits

NPCs generated from hPSCs in mTeSR 1™ using the STEMdiff™ SMADi Neural Induction Kit embryoid body (EB) protocol were differentiated and matured to forebrain-type neurons using the STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits. (A) Forebrain-type neurons were formed after iPS cell-derived NPCs were cultured with the STEMdiff™ Forebrain Neuron Differentiation Kit for 7 days and STEMdiff™ Forebrain Neuron Maturation Kit for 14 days. The resulting cultures contain a highly pure population of (B) class III β-tubulin-positive neurons (green), with (C) fewer than 10% astrocytes (GFAP-positive cells, red). (D) Nuclei are labeled with DAPI (blue).

STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits Are Replacing STEMdiff™ Neuron Kits and Have Equivalent Performance in Deriving Forebrain-type Neurons from Neural Progenitor Cells.

Figure 4. Performance of STEMdiff™ Neuron and the STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits

Neural progenitor cells (NPCs) were derived using the monolayer protocol for the STEMdiff™ SMADi Neural Induction Kit, from H9 ES cells maintained in mTeSR™ 1. NPCs were then differentiated into forebrain-type neurons using either A) STEMdiff™ Neuron Differentiation and Maturation Kits or B) the equivalent-performing STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits according to protocol instructions (see product information sheets for Catalog #08600/08605). The resulting neurons were fixed for immunofluorescence. Both sets of products demonstrate equivalent differentiation efficiency, with a yield of ≥90% class III ꞵ-tubulin-positive neurons (green) and ≤10% GFAP-positive astrocytes (red).

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.

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