STEMdiff™ Astrocyte Differentiation Kit

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

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STEMdiff™ Astrocyte Differentiation Kit

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

1 Kit
Catalog #100-0013
249 USD

Required Products

Overview

STEMdiff™ Astrocyte Differentiation Kit is used to rapidly and efficiently generate astrocytic precursors from neural progenitor cells (NPCs) derived from human pluripotent stem cells (hPSCs) using STEMdiff™ SMADi Neural Induction Kit (Catalog #08581). These astrocytic precursors are then matured further into astrocytes using STEMdiff™ Astrocyte Maturation Kit (Catalog #100-0016). Using this system, a highly pure population of astrocytes (> 70% S100B-positive and > 60% GFAP-positive astrocytes; < 15% doublecortin-positive neurons) can be generated from hPSCs in as few as 7 weeks and can be maintained long term in culture. Cells 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 astrocytes
⦁ Optimized for differentiation from neuronal progenitor cells generated using STEMdiff™ SMADi Neural Induction Kit
⦁ Enables reproducible generation of cortical-type astrocytic precursors derived from multiple human ES and iPS cell lines
Components:
  • STEMdiff™ Astrocyte Differentiation Basal Medium, 80 mL
  • STEMdiff™ Astrocyte 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

Educational Materials

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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

Cortical-type astrocyte precursors can be generated in 20 days from hPSC-derived neural progenitor cells (NPCs) after selecting neural rosettes from replated embryoid bodies. For the maturation of precursors to cortical-type astrocytes, 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 astrocyte precursors can be generated in 21 days 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 cortical-type astrocytes, 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.

PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.