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

STEMdiff™ Forebrain Neuron Differentiation Kit
1 Kit
306 USD
Catalog # 08600

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

Product 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

What's Included

  • STEMdiff™ Forebrain Neuron Differentiation Basal Medium, 80 mL
  • STEMdiff™ Forebrain Neuron Differentiation Supplement, 20 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

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.
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

Data Figures

Figure 1. Schematic for the Embryoid Body Protocol

Forebrain-type neural precursors can be generated in 18 - 19 days from hPSC-derived NPCs after selecting neural rosettes from replated embryoid bodies. For the maturation of precursors to forebrain-type neurons, see the PIS. hPSC = human pluripotent stem cell; NPCs = neural progenitor cells; PIS = product information sheet

Figure 2. Schematic for the Monolayer Protocol

Forebrain-type neural precursors can be generated from 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. NPC = neural progenitor cell; PIS = product information sheet

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 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). NPCs = neural progenitor cells; hPSC = human pluripotent stem cell; EB = embryoid body; iPS = induced pluripotent stem

Figure 4. Downstream Differentiation of Neural Progenitor Cells to Neurons Is Possible Using the STEMdiff™ Differentiation and Maturation Kits

(A) NPCs generated from STiPS-R038 hPSCs in mTeSR™1 using the STEMdiff™ SMADi Neural Induction Kit EB protocol were differentiated and matured to cortical neurons using 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 less than 10% GFAP-positive astrocytes (not shown). (C) The generated neurons are also positive for FOXG1 expression (red), indicating a forebrain-type identity. (D) Nuclei are labeled with Hoechst (blue). NPCs = neural progenitor cells; hPSC = human pluripotent stem cell

Figure 5. A Mixed Population of Forebrain-Type Cortical Neurons Is Generated Using the STEMdiff™ Differentiation and Maturation Kits

Forebrain-type neurons generated from iPSC-derived NPCs (line AIW002-02) were cultured using the STEMdiff™ Forebrain Neuron Differentiation Kit for 7 days and subsequently matured for the following 6 weeks using STEMdiff™ Forebrain Neuron Maturation Kit. The resulting cultures contain a mixed population of neurons expressing VGLUT1, a glutamatergic marker of excitatory neurons (green), as well as MAP2-positive neurons, indicating mature neurons (magenta). Nuclei are labeled with Hoechst (blue). Data courtesy of Cecilia Rocha, The Neuro's Early Drug Discovery Unit (EDDU), McGill University. iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells

Figure 6. 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. NPCs = neural progenitor cells

Figure 7. 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. NPCs = neural progenitor cells

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
Catalog #
08600
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
08600
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
08600
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.

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