In vitro Human Models of Alzheimer´s Disease
Human iPSC-derived (HIP™) neurons from MTI-GlobalStem are seeded, differentiated and matured in 96 well plates for five weeks. The cultures contain 60% neurons and 40% astroglial cells.
Immage above: Neurons derived from human IPS cells, cultured for five weeks and stained for:MAP2 (green) for neurons, GFAP (red) for glia cells, and DAPI (blue) staining cell nuclei.
The expression of the following markers has been verified by Western Blot Analysis: β-tubulin, SNAP25, GFAP, and total tau:
Immages below: The neurons have been verified to express PSD 95 and synaptophysine in a DAPI stained background of nuclei by immunohistochemistry:
The five weeks old human neurons are treated for 24 h with SynAging’s AβO preparation (1 µM based on monomer), with or without BDNF (1.5 nM). The readout indicating neuronal health is a specific marker for neurons: an ELISA for neuron specific enolase (NSE; 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 can be added to this model:
- Gene and protein expression changes by RT-qPCR, immunoblot, or ELISA
- Protein phosphorylation
- Validated ELISA quantification of synaptic markers: e.g. PSD95, Synaptophysin, SNAP25
- Oxidative stress
SynAging’s in vitro human AD assay can investigate three test items, or three test item concentrations, in triplicate, per plate, with the following groups:
- Vehicle (vehicle control)
- AbO (negative control)
- AbO and BDNF / humanin (positive control)
- Test Item in vehicle control
- Test Item in AbO
Please contact us to discuss your interests and requirements for this model.
Product Sheet “In vitroAD Screen on Human Neurons" Download PDF, 550 KB