STEMdiff™ Astrocyte Differentiation Kit
⦁ 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
- STEMdiff™ Astrocyte Differentiation Basal Medium, 80 mL
- STEMdiff™ Astrocyte Differentiation Supplement, 20 mL
Document Type | 产品名称 | Catalog # | Lot # | 语言 |
---|---|---|---|---|
Product Information Sheet | STEMdiff™ Astrocyte Differentiation Kit | 100-0013 | All | English |
Special Protocol | STEMdiff™ Astrocyte Differentiation Kit | 100-0013 | All | English |
Safety Data Sheet 1 | STEMdiff™ Astrocyte Differentiation Kit | 100-0013 | All | English |
Safety Data Sheet 2 | STEMdiff™ Astrocyte Differentiation Kit | 100-0013 | All | English |
Data
![Experimental Protocol Schematic for STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits (Embryoid Body Protocol)](/uploads/2021/100-0013-Fig1.jpg)
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)](/uploads/2021/100-0013-Fig2.jpg)
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](/uploads/2021/100-0013-Fig-3.jpg)
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.
![](/uploads/2021/100-0013-Fig4.jpg)
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.
![](/uploads/2021/100-0013-Fig5.jpg)
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.
![](/uploads/2021/100-0013-Fig6.jpg)
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.