Compounds for use in progressive multiple sclerosis

A method for identifying a compound able to increase oligodendrocyte precursor cell (OPC) differentiation and/or to produce an expanded population of oligodendrocytes, wherein said method comprises:

• a toxicity assay on neonatal mouse oligodendrocyte progenitor cells wherein test compounds are screened for their ability to reduce [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)];
• and/or
• a toxicity assays on rat oligodendrocyte progenitor double fluorescence CG4 line;
• and/or
• a differentiation assay on rat oligodendrocyte progenitor CG4 line.

A method for identifying a compound able to preserve neuronal viability and morphology in cell culture, wherein said method comprises:

• a toxicity assay on primary mouse cortical neurons wherein test compounds are screened in a Cell Counting Kit-8 (CCK-8) assay using WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt);
• and/or
• a neuroprotective assay on primary mouse cortical and striatal neurons wherein compounds are screened for their ability to preserve neuronal viability and morphology (neurite length and network integrity/branching) against NMDA-induced excitotoxicity;
• and/or
• a toxicity assay on iPSC-derived glutamatergic neurons wherein test compounds are screened in a Cell Counting Kit-8 (CCK-8) assay using WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt);
• and/or
• a neuroprotective assay on iPSC-derived glutamatergic neurons wherein compounds are screened for their ability to preserve neuronal viability against ROS (tBuOOH)-induced toxicity.

A method for identifying a compound able to increase neuronal differentiation and/or to produce an expanded population of neurons in cell culture, wherein said method comprises a differentiation assay on human fetal NPCs towards TUJ1+ neuronal precursors wherein:

• test compounds at 1 μM concentration are co-incubated with hufNPCs for 7 days;
• 0.02% vol/vol DMSO and basal medium are used as negative controls;
• heparin is used as a positive control;
• at the end of incubation cells are fixed and analyzed via immunocytochemistry.