Characterization and Chromatographic Isolation of Platelet Extracellular Vesicles from Human Platelet Lysates for Applications in Neuroregenerative Medicine
- Ariunjargal Nyam-ErdeneAriunjargal Nyam-ErdeneInternational PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 101, TaiwanMore by Ariunjargal Nyam-Erdene
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- Ouada NebieOuada NebieGraduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, TaiwanMore by Ouada Nebie
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- Liling DelilaLiling DelilaGraduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, TaiwanMore by Liling Delila
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- Luc BuéeLuc BuéeUniversité de Lille, Inserm U1172, CHU-Lille, Lille Neuroscience & Cognition, Lille 59000, FranceAlzheimer & Tauopathies, Labex DISTALZ, Lille 59000, FranceNeuroTMULille International Laboratory, Université de Lille, Lille 59000, FranceMore by Luc Buée
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- David DevosDavid DevosUniversité de Lille, Inserm U1172, CHU-Lille, Lille Neuroscience & Cognition, Lille 59000, FranceNeuroTMULille International Laboratory, Université de Lille, Lille 59000, FranceMore by David Devos
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- Szu-Yi ChouSzu-Yi ChouPh.D. Program for Neural Regenerative Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei 110, TaiwanNeuroTMULille International Laboratory, Taipei Medical University, Taipei 101, TaiwanMore by Szu-Yi Chou
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- David BlumDavid BlumUniversité de Lille, Inserm U1172, CHU-Lille, Lille Neuroscience & Cognition, Lille 59000, FranceAlzheimer & Tauopathies, Labex DISTALZ, Lille 59000, FranceNeuroTMULille International Laboratory, Université de Lille, Lille 59000, FranceMore by David Blum
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- Thierry Burnouf*Thierry Burnouf*Email: [email protected]. Phone: +886-988-925-235.International PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 101, TaiwanGraduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, TaiwanNeuroTMULille International Laboratory, Taipei Medical University, Taipei 101, TaiwanInternational PhD Program in Cell Therapy and Regeneration Medicine, College of Medicine, Taipei Medical University, Taipei 110, TaiwanBrain and Consciousness Research Centre, TMU Shuang Ho Hospital, New Taipei City 106, TaiwanMore by Thierry Burnouf
Abstract
Human platelet lysates (HPLs) made from clinical-grade platelet concentrates are currently evaluated in the preclinical models of Parkinson’s disease, Alzheimer’s disease, traumatic brain injury, and others, as a new polyvalent neuroprotective biotherapy of the central nervous system. However, the presence and content of extracellular vesicles (EVs) in HPLs and their potential contribution to the neuroprotective and neurorestorative activities of HPLs are still unknown. We, therefore, characterized the EVs present in four different HPL preparations and after purification by size-exclusion chromatography. We then tested the effect of the isolated EVs on neuronal cell repair. We identified that all four HPLs contained a high and similar amount of EVs (1011 to 1012/mL) with a mean size ranging from ca. 50 to 300 nm and a negative zeta potential as determined by nanoparticle tracking analysis and dynamic light scattering. Western blot analysis revealed that the EVs present in HPLs expressed the clusters of differentiation 41 (CD41) and 61 (CD61) characteristic of platelets. These EVs were efficiently isolated from HPL proteins by Sepharose CL-2B size-exclusion column chromatography as confirmed by total protein determination and protein profile by sodium dodecyl sulfate polyacrylamide gel electrophoresis, with 73–85% recovery and maintenance of their size, negative zeta potential, and CD41 and CD61 expression. Interestingly, the EVs purified from the four HPLs exhibited a differential capacity to promote cell growth and migration in a wound-healing assay using SH-SY5Y neuronal cells, and one EV preparation stimulated network formation in primary neuronal cultures. These data indicated that the EVs present in HPLs have different neuroregenerative capacities and that some EV preparations may have interesting applications as a stand-alone therapy for usage in neuroregenerative medicine.
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