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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 a heterozygous (HZ) 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 iCell DopaNeurons SNCA A53T HZ. Please also see the isogenic iCell DopaNeurons.
iCell DopaNeurons SNCA A53T HZ exhibit the relevant biology and functionality to advance research and preclinical studies for devastating neurological disorders:
qPCR analysis reveals decreased TH and DDC and increased COMT expression in A53T dopaminergic neurons suggesting that biosynthetic levels of DA in the SNCA A53T neurons should be decreased.
Immunofluorescence staining shows detectable levels of alpha-synuclein in the control cells and more in the SNCA A53T DA neurons (data not quantified).
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.
Both iCell DopaNeurons and iCell DopaNeurons SNCA A53T HZ display high activity and synchronous bursting on MEA; A53T cells have fewer bursts per minute, but the intensity of those bursts are ~5X greater.
iCell SNCA A53T HZ 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.
iCell DopaNeurons are shipped cryopreserved with optimized media. Simply thaw and use.
iCell DopaNeurons are fully differentiated neurons, not precursors, providing biologically relevant and reproducible results.
iCell DopaNeurons are >80% tyrosine hydroxylase positive (THpos) midbrain dopaminergic neurons, enabling mechanistic studies of neurodegenerative diseases.
iCell DopaNeurons are floor plate-derived dopaminergic neurons from human iPS cells ensuring complete regional specification for full functionality.