STEMdiff™ Forebrain Neuron Maturation Kit

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

STEMdiff™ Forebrain Neuron Maturation Kit

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

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Maturation kit for generation of functional neurons from human ES and iPS cell-derived neuronal precursor cells
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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 maturation of neuronal precursors generated using STEMdiff™ Forebrain Neuron Differentiation 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

  • BrainPhys™ Neuronal Medium, 100 mL
  • STEMdiff™ Forebrain Neuron Maturation Supplement, 25 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

The STEMdiff™ Forebrain Neuron Maturation Kit is used in conjunction with the STEMdiff™ Forebrain Neuron Differentiation Kit (Catalog #08600) to generate a mixed population of forebrain-type (FOXG1-positive) neurons from neural progenitor cells derived from human pluripotent stem cells. The neurons derived are highly pure (≥ 90% class III β-tubulin-positive neurons; < 10% GFAP-positive astrocytes), functional and can be maintained long-term in culture. 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, Neurons
Species
Human
Application
Cell Culture, Characterization, Differentiation, Functional Assay, Immunocytochemistry, Phenotyping
Brand
STEMdiff
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Neuroscience
Formulation Category
Chemically Defined, Serum-Free

Data Figures

Figure 1. Schematic for the Embryoid Body Protocol

Forebrain-type neurons can be generated from NPCs after 6 days in STEMdiff™ Forebrain Neuron Differentiation Medium. For differentiation of precursors from embryonic and induced pluripotent stem cells, see the PIS. NPCs = neural progenitor cells; PIS = product information sheet

Figure 2. Schematic for the Monolayer Protocol

Forebrain-type neurons can be generated from NPCs after 6 days in STEMdiff™ Forebrain Neuron Differentiation Medium. For differentiation of precursors from embryonic and induced pluripotent stem cells, see the PIS. NPC = neural progenitor cells; 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 #
08605
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
08605
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
08605
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.