Neural Stem Cells

Control-iPSCs     iPSC-NSC-astrocytes     iPSC-NSC-dopaminergicneurons     iPSC-NSC-neuron-mixed     iPSC-NSC-differentiation-induction-media                   

Neural Stem Cells (NSC) are multipotent cells derived from iPSCs and ESCs that are self-renewing, and have potential to differentiate into various neuronal lineage cells. This makes them very attractive for in vitro patient-specific neuroscience research, whereby NSCs can be derived from patient-specific PSCs and further differentiated into CNS neurons and glial cells.

Applied StemCell offers high quality neural stem cells derived from fully characterized iPSC lines from multiple donors, for flexibility in choosing the lineage most appropriate for your research. These NSCs have been derived using integration-free, proprietary neural induction protocols, express neural stem cell markers such as PAX6, SOX1 and nestin and form neural rosettes. The NSCs retain their multipotency and neural markers even after cryopreservation and passaging.  These fully characterized NSCs at low passage can be further differentiated to the particular neuronal cell type of the investigator’s choice, thus facilitating studies in cell replacement therapies and neuronal disease modeling. 

SingleiPSC-differentitiaion-NSC&derivatives


Advantages of choosing ASC’s iPSC-derived neural stem cells:

  • Different NSC lines, each derived from a single control iPSC line or engineered iPSC line: male (cord blood), female (cord blood), male (fibroblasts) and more
  • High purity (>90%) cells expressing neural stem cell markers: SOX1, PAX6 and Nestin
  • Fully characterized by immunocytochemistry and whole genome profiling
  • Provides a consistent and reliable source of differentiated mature, functional neurons and glial cells
  • Physiologically relevant cell line models

Applications:

  • Neural development
  • Neurotoxicity
  • Gene Profiling during differentiation
  • Disease Modeling
  • Electrophysiology
  • Co-culture applications
Also available: Isogenic dopaminergic neurons, astrocytes, and other CNS neurons from the each control iPSC line. 
 
 
TECHNICAL-NSC-Nestin-Sox1-DAPI-2
 
Figure 1. Neural Stem Cells (ASE-9234) derived from a male cord blood iPSC line, is a homogenous population of cells expressing > 98% Nestin (green), > 90% Sox1 (Red) and DAPI (blue). This cell line has been used for differentiation into astrocytes, dopaminergic neurons and CNS neurons, and extensively for neurotoxicity and neuroprotection screening assays.
 
NSC differentiation Figure 1 image - Neural Stem Cell Differentiation

Figure 2. Neural stem cells (ASE-9303) differentiated from a well-characterized control fibroblast-derived iPSC line (ASE-9203) using Applied StemCell's proprietary neural induction protocol and NSC culture media. Applied StemCell’s differentiated NSCs retain neural stem cell phenotype even after passaging and freeze-thaw under feeder-free conditions as evidenced by staining for NSC markers PAX6 (>90%) and SOX1 (>90%) and nuclear DNA (DAPI). Note: White arrows indicate distinctive neuronal rosettes. 

 

Neural Stem Cell Differentiation Services: We can differentiate your control/ patient-derived/ engineered iPSCs into nerual stem cells and further into neurons and glial cells, including full characterization of your derived cell lines.


Technical Details

Characterization of neurons derived from neural stem cells (ASE-9234)

TECHNICAL-NSC-neuroncharacterization

Figure 1. Expression of neuronal markers in neurons differentiated from NSC from male cord blood iPSCs (ASE-9234). Immunocytochemical characterization of neurons differentiated from NSC shows high expression of Tuj1, MAP2 and GABA neuronal markers and cells. These NSCs can therefore be differentiated to cells of a specific lineage with high purity and functionality.

 TECHNICAL-neuron-wholegenomeprofiling

Figure 2. Whole genome profiling for markers expressed by neurons derived from NSCs.


Charaterization of astrocytes derived from neural stem cells (ASE-9234)

TECHNICAL-Astrocytes-GFAPstaining

Figure 1. Immunocytochemical characterization of astrocytes derived from NSCs (ASE-9234). Astrocyte marker GFAP (Red) is expressed in > 90% of the cells while neuronal marker Tuj1 is expressed in <1% of the cells; (data not shown).

TECHNICAL-astrocyte-wholegenomeprofiling

Figure 2. Whole genome profiling for markers expressed by astrocytes derived from NSCs.


Co-culturing of neurons and astrocytes to develop complex models

Neurons and astrocytes derived from Applied StemCell's NSCs can be co-cultured for developing complex research models. The cells can be isogenic differentiated cells or cells from different genetic backgrounds.

TECHNICAL-Astrocyte&neurons-coculture

Figure. Enhanced synpase formation in neuron-astrocyte co-cultures. Co-culture of neurons and astrocytes showed a significant increase in synaptic puncta, as seen by the co-colocalization of neuronal marker (Tuj1) and synapse marker (Synapsin),as compared to neuron only cultures.  

References

Shaltouki, A., Sivapatham, R., Pei, Y., Gerencser, A. A., Momčilović, O., Rao, M. S., & Zeng, X. (2015). Mitochondrial alterations by PARKIN in dopaminergic neurons using PARK2 patient-specific and PARK2 knockout isogenic iPSC lines. Stem cell reports4(5), 847-859.

Efthymiou, A. G., Steiner, J., Pavan, W. J., Wincovitch, S., Larson, D. M., Porter, F. D., ... & Malik, N. (2015). Rescue of an in vitro neuron phenotype identified in Niemann-Pick disease, type C1 induced pluripotent stem cell-derived neurons by modulating the WNT pathway and calcium signaling. Stem cells translational medicine4(3), 230-238.

Efthymiou, A., Shaltouki, A., Steiner, J. P., Jha, B., Heman-Ackah, S. M., Swistowski, A., ... & Malik, N. (2014). Functional screening assays with neurons generated from pluripotent stem cell–derived neural stem cells. Journal of biomolecular screening19(1), 32-43.

Shaltouki, A., Peng, J., Liu, Q., Rao, M. S., & Zeng, X. (2013). Efficient generation of astrocytes from human pluripotent stem cells in defined conditions. Stem cells31(5), 941-952.


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