A Novel Ex Vivo Model to Study Therapeutic Treatments for Myelin Repair following Ischemic Damage.
Luisa WernerMichael GliemNicole RychlikGoran PavicLaura ReicheFrank KirchhoffMarkley Silva Oliveira JuniorJoel GruchotSven G MeuthPatrick KüryPeter GöttlePublished in: International journal of molecular sciences (2023)
Stroke is a major reason for persistent disability due to insufficient treatment strategies beyond reperfusion, leading to oligodendrocyte death and axon demyelination, persistent inflammation and astrogliosis in peri-infarct areas. After injury, oligodendroglial precursor cells (OPCs) have been shown to compensate for myelin loss and prevent axonal loss through the replacement of lost oligodendrocytes, an inefficient process leaving axons chronically demyelinated. Phenotypic screening approaches in demyelinating paradigms revealed substances that promote myelin repair. We established an ex vivo adult organotypic coronal slice culture (OCSC) system to study repair after stroke in a resource-efficient way. Post-photothrombotic OCSCs can be manipulated for 8 d by exposure to pharmacologically active substances testing remyelination activity. OCSCs were isolated from a NG2-CreERT2-td-Tomato knock-in transgenic mouse line to analyze oligodendroglial fate/differentiation and kinetics. Parbendazole boosted differentiation of NG2 + cells and stabilized oligodendroglial fate reflected by altered expression of associated markers PDGFR-α, CC1, BCAS1 and Sox10 and GFAP. In vitro scratch assay and chemical ischemia confirmed the observed effects upon parbendazole treatment. Adult OCSCs represent a fast, reproducible, and quantifiable model to study OPC differentiation competence after stroke. Pharmacological stimulation by means of parbendazole promoted OPC differentiation.
Keyphrases
- induced apoptosis
- oxidative stress
- acute myocardial infarction
- stem cells
- multiple sclerosis
- poor prognosis
- atrial fibrillation
- drinking water
- high throughput
- white matter
- young adults
- left ventricular
- magnetic resonance
- cerebral ischemia
- cell proliferation
- long non coding rna
- binding protein
- coronary artery disease
- brain injury
- signaling pathway
- ischemia reperfusion injury