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Whole and fractionated human platelet lysate biomaterials-based biotherapy induces strong neuroprotection in experimental m ... 1 1 Introduction 1 2 Material and methods 2 2.1 HHPL and fractions 2 2.1.1 Preparation of heat-treated human platelet lysate (HHPL) and fractions 2 2.1.2 HHPL and fractions characterization 2 2.2 LUHMES culture 2 2.2.1 Cell maintenance and differentiation 2 2.2.2 Cell treatment and viability assay 2 2.2.3 siRNA transfection 3 2.3 Primary motoneuron culture 3 2.3.1 Motoneuron isolation 3 2.3.2 Cell treatment and survival assay 3 2.4 Animal experiments 3 2.4.1 Mice 3 2.4.2 Riluzole drug administration 3 2.4.3 Intracerebroventricular (I.C.V) injection and intranasal administration (I.N) 3 2.4.4 RNA extraction and RT-qPCR for acetylcholine receptor alpha (AchRα) 3 2.4.5 Neurofilament light chain (NfL) quantification 3 2.5 Statistical analysis 3 3 Results 3 3.1 Fractions of HHPL protect dopaminergic neurons against cell death 3 3.2 Neuroprotective pathways elicited by HHPL and the <3 kDa fraction against ferroptosis differentially rely on GPX4 5 3.3 HHPL but not the <3 kDa fraction protects primary motor neurons against excitotoxic, apoptotic and oxidative in

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nonfiction