Applied StemCell's provides cryo-preserved, pre-differentiated mixed population neuronal precursors derived from a footprint-free, karyotype normal human iPSC line. They yield high purity, functional neurons within 8 days when using Applied StemCell’s well-optimized maturation medium and supplements. The derived-neurons have also been tested for neuronal activity using functional and screening assays. The neurons can be seeded on various culture vessel formats including 96-well plates on either glass or plastic surfaces and cultured as adherent cells. Shortly after seeding, the cells proliferate slightly for up to 3 days and show extensive neurite outgrowth and proper neuronal morphology. In general, on Day 8 post-seeding, the cell population will contain >90% neurons and < 5% Glial Fibrillary Acidic Protein (GFAP) positive astrocytes.
Advantages of Applied StemCell's ready-to-use neurons:
- High purity neurons: >90% Tuj1+ neurons and < 5% GFAP+ cells
- High cell viability of cyropreserved neurons (>80%)
- Available as isogenic lineages of mixed neurons and associated neuronal derivatives from two different control iPSC lines (male ASE-9109 and female ASE-9110)
- Functionally viable neurons capable of neuronal excitability and synapse formation
- Long-term viability in cell culture and consistency in results (>30 days): affords greater flexibility in designing high throughput drug screening and testing
- Electrophysiology and synaptic functionality assays
- Neurotoxicity and neuroprotection screening tests
- Physiologically relevant disease modeling platform
- Gene profiling under isogenic conditions to understand gene functionality associated with neurological diseases
Also available: Isogenic dopaminergic neurons, and astrocytes from the each control iPSC line. Applied StemCell's catalog also provides Neuron Starter Kit to generate and maintain mature, functional neurons in your own lab.. The kits include cryopreserved pre-differentiated neurons as well as optimized medium and supplements.
Figure 1. Immunocytochemical characterization of neurons derived from male cord blood control iPSC line (ASE-9109). Immunocytochemical staining of neurons differentiated from iPSC indicate > 90% of the cells stain positive for neuronal markers Tuj1 (green) and GABA (red); < 5% are GFAP+ cells. indicating high purity of neuronal cell population.
Figure 2. Whole genome profiling for markers expressed by neurons derived from NSCs.
Custom Differentiation Service for iPSC-derived Neurons: We can direct differentiaion of your control/ patient/ engineered iPSC lines into functional cortical/ motor neuron lineages, including full characterization of the derived cell lines.
Isogenic neural stem cell derivatives differentiated from a single iPSC line
Applied StemCell's Neural Stem Cells (NSCs) can be differentiated to neuron products including neurons and glial cells (dopaminergic and mixed neurons, astrocytes and oligodendrocytes) using ASC's differentiation and maturation media and kits. NSCs used for further neural lineage cell derivation uniformly express >90% Sox1 and >98% Nestin. These neural stem cell-derived neurons and astrocytes have been used extensively for screening assays in publications.
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.
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.
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 reports, 4(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 medicine, 4(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 screening, 19(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 cells, 31(5), 941-952.