Dopaminergic Neurons

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SCHEMATIC-singleiPSC-differentiation-neuronallineage-catalog#s

For several years, neuroscience research using cell line and animal models of degenerative neurological disorders such as Parkinson’s disease (PD) and Alzheimer’s has been plagued with the lack of translation of results to the human scenario, whether for basic understanding of disease pathology or for screening of targeted drug therapies. The development of the induced pluripotent stem cell (iPSC) technology and its further differentiation into various post-mitotic somatic cell lineages, especially neuronal lineage cells and dopaminergic neurons, has overcome a major bottleneck by providing biologically relevant in vitro models to screen drugs for neuroprotection and movement-disorder therapy.

Applied StemCell's Human Dopaminergic neurons are cryo-preserved, pre-differentiated dopaminergic neuron precursors derived from a footprint-free, karyotype normal human iPSC line. These precursors yield high purity mature dopaminergic neurons using Applied StemCell’s optimized dopaminergic maturation medium and supplements (ASE-9323DM). Mature dopaminergic neurons can be obtained within 12 days after thaw, and the matured dopaminergic neurons can be maintained in culture for up to 3 weeks without losing functionality.  These mature and functional dopaminergic neurons contain >80% neurons, >30% TH+ dopaminergic neurons and < 15% GFAP+ astrocytes.

Advantages of using ASC’s iPSC-derived Dopaminergic neurons:

  • Yields high purity, functional dopamine neurons after maturation
  • Mature dopamine neurons can be maintained long-term in culture (up to 3 weeks)
  • Differentiated from NSCs using footprint-free, integration-free protocols
  • Two dopaminergic lineages (ASE-9323, ASE-9323F): derived from one male donor (ASE-9109) and one female donor (ASE-9110)
  • Also, available: isogenic panels of neural stem cells, other differentiated neurons, astrocytes precursors and mature cells from same donor iPSCs
  • Proven as an ideal model for disease modeling, neuroprotection and neurotoxicity screening assays

Applications:

  • Reliable predictive models of neurological disorders
  • Drug screening
  • Neuroprotection assays and research
  • Neurotoxicity screening
Also available: Isogenic mixed population of neurons, and astrocytes from the each control iPSC line.  Applied StemCell's catalog also provides Dopaminergic Neuron Starter Kit to differentiate and maintain mature, functional neurons in your own lab.. The kits include cryopreserved dopamine neuron precursor cells as well as optimized medium and supplements .
 

TECHNICAL-DA-neurons

Figure 1. Immunocytochemical characterization of mature dopaminergic neurons (ASE-9323) differentiated from iPSC-derived neural stem cells (parent iPSC line: ASE-9109). Top left: Bright field image indicates healthy dopaminergic cell morphology at 12 days post-seeding. Other panels: Immunostaining on day 12 post-seeding shows that >90% of total cells expressed Tuj-1 marker (green) and >30% expressed TH marker (red). A count of the number of nuclei (DAPI; blue) was used to calculate the total number of cells. 

 

Custom Differentiation Service for iPSC-derived Dopaminergic Neurons: We can direct differentiaion of your control/ patient/ engineered iPSC lines into functional dopaminergic neurons, including full characterization of the cell lines.

Technical Details

Isogenic neural stem cell derivatives differentiated from a single iPSC line


TECHNICAL-singleiPSC-differentiation-neuronallineage

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.

 TECHNICAL-Astrocyte-Neuron-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 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.

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