- 作者: Ouada Nebie, David Devos, Valérie Vingtdeux, Lassina Barro, Jean-Christophe Devedjian, Aurélie Jonneaux, Ming-Li Chou, Régis Bordet, Luc Buée, Folke Knutson, David Blum and Thierry Burnouf
- 作者服務機構: 1. Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, 250 Wu-Xing Street, Taipei, 11031, Taiwan 2. Univ Lille, Inserm, CHU Lille, UMR-S1171. Lille Neuroscience & Cognition, Degenerative and vascular cognitive disorders, F-59000, Lille, France 3. David Devos and Valérie Vingtdeux contributed equally to this work. 4. Univ. Lille, Inserm, CHU-Lille, UMR-S1172, Lille Neuroscience & Cognition, Alzheimer & Tauopathies, F-59000, Lille, France 5. International Ph.D. Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan 6. Present address: INSERM UMRS 938, CdR Saint-Antoine, Laboratory Immune System, Neuroinflammation and Neurodegenerative Diseases, Saint-Antoine Hospital, Paris, France 7. Clinical Immunology and Transfusion Medicine IGP, Uppsala University, Uppsala, Sweden 8. International Ph.D. Program in Cell Therapy and Regeneration Medicine, Taipei Medical University, Taipei, Taiwan
- 中文摘要:
- 英文摘要:
Background
Effective neurorestorative therapies of neurodegenerative diseases must be developed. There is increasing interest in using human platelet lysates, rich in neurotrophic factors, as novel disease-modifying strategy of neurodegeneration. To ensure virus safety, pathogen reduction treatments should be incorporated in the preparation process of the platelet concentrates used as source material. We therefore investigated whether platelet concentrates (PC) pathogen-inactivated using a licensed photo-inactivation treatment combining photosensitive psoralen (amotosalen) and UVA irradiation (Intercept) can serve as source material to prepare platelet lysates with preserved neuroprotective activity in Parkinson’s disease models.
Methods
Intercept treated-PCs were centrifuged, when reaching expiry day (7 days after collection), to remove plasma and platelet additive solution. The platelet pellet was re-suspended and concentrated in phosphate buffer saline, subjected to 3 freeze-thaw cycles (− 80 °C/37 °C) then centrifuged to remove cell debris. The supernatant was recovered and further purified, or not, by heat-treatment as in our previous investigations. The content in proteins and neurotrophic factors was determined and the toxicity and neuroprotective activity of the platelet lysates towards LUHMES cells or primary cortical/hippocampal neurons were assessed using ELISA, flow cytometry, cell viability and cytotoxicity assays and proteins analysis by Western blot.
Results
Platelet lysates contained the expected level of total proteins (ca. 7–14 mg/mL) and neurotrophic factors. Virally inactivated and heat-treated platelet lysates did not exert detectable toxic effects on neither Lund human mesencephalic dopaminergic LUHMES cell line nor primary neurons. When used at doses of 5 and 0.5%, they enhanced the expression of tyrosine hydroxylase and neuron-specific enolase in LUHMES cells and did not significantly impact synaptic protein expression in primary neurons, respectively. Furthermore, virally-inactivated platelet lysates tested were found to exert very strong neuroprotection effects on both LUHMES and primary neurons exposed to erastin, an inducer of ferroptosis cell death.
Conclusion
Outdated Intercept pathogen-reduced platelet concentrates can be used to prepare safe and highly neuroprotective human heat-treated platelet pellet lysates. These data open reassuring perspectives in the possibility to develop an effective biotherapy using virally-inactivated platelet lysates rich in functional neurotrophins for neuroregenerative medicine, and for further bio-industrial development. However, the data should be confirmed in animal models. - 中文關鍵字:
- 英文關鍵字: Pathogen inactivation, Intercept-platelet lysate, Ferroptosis, Neuroprotection, LUHMES cells, Primary neurons, Synaptic markers