Applied StemCell’s Dopaminergic neurons are derived from directed differentiation of well-characterized control human iPSC lines using integration-free, footprint-free differentiation protocols. These iPSC-derived dopaminergic neurons are high purity, functional dopamine neurons. They are provided as dopamine precursor cells that can be matured to primary dopamine neurons using an easy maturation protocol, and can be maintained in culture for up to 3 weeks. These neurons are excellent as experimental controls for research with dopamine neurons derived from disease iPSC lines, for co-culture modeling, drug and neurotoxicity screening applications.
No, the ASE-9323 dopaminergic precursors are not suitable for expansion and cryopreservation once seeded. Once they mature into dopaminergic neurons, we recommend their use for only up to 3 weeks.
The ASE-9323K Dopaminergic neuron kit comes with 1 vial of dopamine neuron precursor cells, DOPA maturation medium, and supplements to aid the maturation of dopaminergic neurons. The ASE-9323 Dopaminergic neuron precursor cells contain only 1 vial of precursor cells and do not come with media and supplements. The maturation media and supplement (ASE-9323DM) are available for purchase separately.
The cells are post-mitotic and the yield can vary depending on the person handling the cells.
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 synapse 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.
Designing co-culture assays: Comparing timelines for astrocytes, neurons and DOPA neurons derived from neural stem cell (ASE-9234)
Figure. Chart detailing the differentiation and maturation timelines of neural stem cells (ASE-9234), astrocytes (ASE-9322), neurons (ASE-9321), and DOPA neurons (ASE-9323) to design co-culture assays.
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.
ASC’s Human iPSC-derived Dopaminergic Neurons are provided as late stage dopamine neuron precursors derived from footprint-free, karyotype normal human iPSC lines. Using an straightforward, maturation protocol, mature and functional dopaminergic neurons can be obtained that contain >80% Tuj1+ neurons and >30% TH+ dopaminergic neurons, and can be used for long-term culture.
Advantages of using ASC’s iPSC-derived Dopaminergic neurons:
- Dopamine neurons that are positive for dopamine marker and capable of functional dopamine-release
- Differentiated from NSCs using footprint-free, integration-free protocols
- Two dopaminergic lineages each derived from one male donor (ASE-9109) and one female donor (ASE-9110)
- Matched, 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, co-culture assays, neuroprotection and neurotoxicity screening assays
- Controls for predictive models of neurological disorders
- Drug screening
- Neuroprotection assays and research
- Neurotoxicity screening
Also available: Isogenic mixed 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.
Figure 1. Immunocytochemical characterization of control dopaminergic/dopamine neurons (ASE-9323) at day 22 of differentiation from iPSC-derived neural stem cells (parent iPSC line: ASE-9109). Maturation of dopaminergic neurons can be assessed by their morphology and by immunostaining using three markers. Percentage of dopaminergic neurons can be determined by TH (Tyrosine Hydroxylase) positive neurons divided by the total number of cells (DAPI staining of nuclei). Green: Tuj1 (neuronal class III b-Tubulin; >80%); Red: TH; Blue: DAPI.
Custom Differentiation Service for iPSC-derived Dopaminergic Neurons: Applied StemCell provides directed differentiation of your control/ patient/ engineered iPSC lines into functional dopaminergic neurons, including full characterization of the cell lines.
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.