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MyCell® DopaNeurons (A53T)

Study a Highly Penetrant Mutation in Human Dopaminergic Neurons

SNCA A53T is one of the most highly penetrant and widely studied mutations linked to Parkinson’s disease (PD). SNCA encodes for the alpha-synuclein (α-syn) protein, which is predominantly expressed in the brain at presynaptic terminals. The A53T mutation renders α-syn more susceptible to aggregation and accumulation, which are hallmark indicators of PD pathology.

To generate an A53T allelic variant isogenic to iCell® DopaNeurons, nuclease-mediated SNP alteration of a healthy control iPS cell line was performed to introduce this site-specific mutation into the gene for SNCA. This genome-engineered iPS cell was then differentiated into human midbrain floorplate dopaminergic (DA) neurons according to protocols licensed and adapted from the Lorenz Studer lab (Memorial Sloan Kettering) and industrialized at CDI, to produce MyCell® DopaNeurons (A53T). Please also see the isogenic iCell DopaNeurons.

Best-in-Class Biology

MyCell DopaNeurons (A53T) exhibit the relevant biology and functionality to advance research and preclinical studies for devastating neurological disorders:

  • Fully differentiated, >80% pure midbrain DA neurons
  • Expression of relevant midbrain DA neuron markers (e.g. Lmx1, FoxA2, and TH)

Comparison to Isogenic iCell DopaNeurons

Figure 1: qPCR analysis reveals decreased TH and DDC and increased COMT expression in A53T dopaminergic neurons suggesting that biosynthetic levels of dopamine in the SNCA A53T neurons should be decreased.

Figure 2: Immunofluorescence staining shows detectable levels of alpha-synuclein in the control cells and more in the SNCA A53T DA neurons (data not quantified).

Figure 3: Bioenergetic analysis of WT versus A53T cells indicates that the response to FCCP and the max respiratory capacity is greater in the mutant line. Data are normalized based on cell number.

Figure 4: Both iCell DopaNeurons and MyCell DopaNeurons (A53T) display high activity and synchronous bursting on MEA; A53T cells have fewer bursts per minute, but the intensity of those bursts are ~5X greater.

Figure 5: MyCell SNCA A53T DopaNeurons display spontaneous Ca2+ oscillations earlier in culture (e.g. day 7) with an amplitude and bursting pattern that is significantly different than the WT control.

 

 

  • Ordering Information
    Catalog # Description Quantity
    M1029 iCell Neural Supplement B

    Dopaminergic neurons medium supplement, serum-free

    2 ml Request Quote
    M1031 iCell Nervous System Supplement

    Growth factor supplement for neuronal cell types

    1 ml Request Quote
    01279.431 DDP-DNC-0.5X MyCell DopaNeurons (A53T) Kit

    Each kit contains:

    • Cryopreserved MyCell DopaNeurons (A53T)
    • 100 ml iCell DopaNeurons Maintenance Medium
    • 2 ml iCell DopaNeurons Medium Supplement
    • 1 ml iCell Nervous System Supplement
    ≥2.5 x 106
    Viable Cells/Vial
    Request Quote