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iCell Microglia PD (SNCA)

Alpha-synuclein A53T PD Model with an Isogenic Control

Description

Parkinson’s disease (PD) affects ~1% of people over the age of 65 and is the second most common neurodegenerative brain disorder. Αlpha-synuclein (encoded by the SNCA gene) is a 140 amino-acid protein, expressed highly in dopaminergic neurons of the substantia nigra pars compacta and is intracellularly localized in presynaptic terminals. Alpha-synuclein proteins have the capacity to self-assemble, from unfolded monomers to oligomeric species, to heavy aggregates (called amyloid fibrils).

SNCA (A53T) Mutation Site

Figure 1. Details of the SNCA Mutation
An engineered base change associated with the SNP rs104893877 results in a change of amino acid residue 53 from alanine to threonine.

Accumulation of these insoluble fibrils progressively promotes the formation of intracellular inclusions called Lewy bodies within neurons and glial cells. Recent studies suggest that alpha-synuclein oligomers bind to lipids, disrupt cellular membrane integrity, and induce microglial activation leading to the death of dopaminergic neurons.

Engineered SNCA (A53T) Model

A known mutation in the N-terminal amphipathic domain of alpha-synuclein has been linked to familial Parkinson’s disease. FCDI created a model of this mutation in the iCell® Microglia genetic background line, 01279. The mutation results in a single amino acid change at aa53 from alanine to threonine (Fig. 1).

Characterization

Figure 2. Comparative Marker Expression of SNCA A53T

Comparative Marker Expression
iCell Microglia were labeled for the presence of cell surface (CD45, CD11b, CD11c, TREM2 and, CD33) and intracellular (P2RY12, TMEM119, CX3CR1, IBA1) antigens by flow cytometry (Fig. 2). The specific staining of the engineered iCell Microglia SNCA A53T is compared against matched isotype control.

Phagocytosis
In Figure 3, a comparison is shown between iCell Microglia and the engineered SNCA A53T line. Phagocytosis of S.aureus BioParticles® was measured over time using an IncuCyte® S3 Live-Cell Analysis System. The engineered microglia exhibit phagocytic activity, but at a reduced activity relative to the unengineered line.

Cytokine Response
The cytokine response of stimulated iCell Microglia and the engineered SNCA line were compared by multiplex assay (Fig. 4). iCell Microglia were plated in microglia maintenance media and allowed to recover for 72 h prior to stimulation with LPS or interferon gamma or a combination of both for 24 h. Supernatants were collected and assayed using a Luminex® multiplex cytokine assay.

Figure 3. Comparison of Phagocytosis Activity of SNCA A53T to the Unmodified Line

Figure 4. Cytokine Comparison of iCell Microglia and SNCA

  • Ordering InformationFor best pricing please log in or register
    Catalog # Description Quantity Price
    R1200 iCell Microglia Kit, SNCA (A53T), 01279

    Each kit contains:

    • C1133 Cryopreserved iCell Microglia SNCA (A53T), 01279
    • M1034 50ml iCell Glial Base Medium
    • M1036 0.5ml iCell Microglia Supplement A (100x)
    • M1037 0.5ml iCell Microglia Supplement B (100x)
    • M1035 2 x 1ml iCell Neural Supplement C (25X)
    • X1022 iCell Microglia Quick Guide
    • Additional reagents required; see Quick Guide
    ≥1 x 106
    Viable Cells/Vial
    $2275
    Inquire About Pricing & Availability
    C1133 iCell Microglia, SNCA (A53T), 01279

    Product contains:

    • C1133 Cryopreserved iCell Microglia SNCA (A53T), 01279
    • X1022 iCell Microglia Quick Guide
    • Additional reagents required; see Quick Guide
    ≥1 x 106
    Viable Cells/Vial
    $1975
    Inquire About Pricing & Availability
    R1131 iCell Microglia Kit, 01279

    Each kit contains:

    • C1110 Cryopreserved iCell Microglia, 01279
    • M1034 50ml iCell Glial Base Medium
    • M1036 0.5ml iCell Microglia Supplement A (100x)
    • M1037 0.5ml iCell Microglia Supplement B (100x)
    • M1035 2 x 1ml iCell Neural Supplement C (25X)
    • X1022 iCell Microglia Quick Guide
    • Additional reagents required; see Quick Guide
    ≥1 x 106
    Viable Cells/Vial
    $1650
    Inquire About Pricing & Availability
    C1110 iCell Microglia, 01279

    Product contains:

    • C1110 Cryopreserved iCell Microglia, 01279
    • X1022 iCell Microglia Quick Guide
    • Additional reagents required; see Quick Guide
    ≥1 x 106
    Viable Cells/Vial
    $1350
    Inquire About Pricing & Availability
    R1204 iCell Microglia Media Kit

    Each kit contains:

    • M1034 50ml iCell Glial Base Medium
    • M1036 0.5ml iCell Microglia Supplement A (100x)
    • M1037 0.5ml iCell Microglia Supplement B (100x)
    • M1035 2 x 1ml iCell Neural Supplement C (25X)
    • X1022 iCell Microglia Quick Guide
    1 Kit $365
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    019-19741 Anti IBA1 Polyclonal Antibody
    • Rabbit polyclonal IgG
    • 50μg supplied at 0.5mg/ml in TBS
    • Protocol included
    • Store at -20° C
    50ug $355
    Inquire About Pricing & Availability