In vitro Human Models of Neurodegeneration in Parkinson´s Disease
Human iPSC-derived (HIP™) neurons from Thermo Fisher (before MTI-GlobalStem) are seeded, differentiated for five weeks and matured in 96 well plates for five weeks. The cultures contain 60% neurons and 40% astroglial cells.
The expression of the following markers has been verified by Western blot analysis: β-tubulin, SNAP25, GFAP, and total tau (see below).The neurons have been verified to express PSD95 and synaptophysine in a DAPI stained background of nuclei by immunohistochemistry (see: human AD in vitro model).
The presence of dopaminergic neurons in the human neuronal culture has been verified by Western blot for dopamine transporter (DAT), tyrosine hydroxylase (TH), and α-synuclein:
The 5 weeks matured human neuronal cultures are treated for 72 h with SynAging’s human α-synuclein oligomer (aSO) preparation (1 µM, based on monomer), with or without BDNF. The readout indicating neuronal survival in this mixed culture of neurons and astroglia is a specific marker for neurons: ELISA for neuron specific enolase (NSE, see reference below).
(Ref.: Oliva D, Calì L, Feo S, Giallongo A (1991). "Complete structure of the human gene encoding neuron-specific enolase.". Genomics 10 (1): 157–65)
Further read-outs are available for this model:
- Gene and protein expression changes by RT-qPCR, immunoblot, or ELISA
- Protein phosphorylation
- Validated ELISA quantification of synaptic markers from neurons: e.g. PSD95, Synaptophysin, SNAP25
- Oxidative Stress
SynAging’s in vitro human PD assay can investigate three test items, or three test item concentrations, per plate in triplicate with the following groups:
- Vehicle (vehicle control)
- aSO (negative control)
- aSO + BDNF (positive control)
- Test Item + vehicle
- Test Item + aSO
Please contact us to discuss your interests and requirements for this model.
Product Sheet “In vitro PD Screen on Human Neurons” Download PDF, 550 KB