Characterisation of the Lipidome of Transfusible Platelet Components
- Publication Type:
- Thesis
- Issue Date:
- 2022
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Conventional room temperature stored platelets have a short shelf life and require constant agitation. As such, there can be issues when supplying these components to austere locations. Alternative storage techniques, such as cold storage (2-6°C) and cryopreservation (-80C with DMSO), represent feasible options to circumvent these issues. Extensive efforts have been undertaken to understand the changes occurring in alternatively stored platelet components. Historic studies have assessed the lipidome of cold-stored platelets, however, an updated assessment was warranted due to technological advancements. Further, the lipidome of cryopreserved platelets had not yet been characterised. Therefore, the aim of this thesis was to characterise the changes occurring to the lipid profile of alternatively stored platelet components. Apheresis derived platelet components were stored at either room temperature (20-24C with constant agitation) or cold-stored (2-6°C without agitation) and sampled on day 1, 5, and 14 post-collection. Buffy coat derived platelet components were frozen (-80°C) with 5-6% DMSO, thawed and then stored at room temperature for 24 hours, and samples were taken before freezing, after thawing and after post-thaw storage. The platelet, microparticle and supernatant fractions were separated by differential centrifugation. The lipid profile of the component fractions were assessed by LC-MS/MS. The lipid profile of platelets was relatively unchanged during storage for 5 days, regardless of temperature. However, over extended storage changes became apparent, and these were exaggerated by cold storage. Conversely, the lipid profile of the supernatant was changed during early storage, but changes stabilised during extended storage. Specifically, the proportion of the procoagulant lipids, PS and PE, increased during extended cold storage. Further, several LPC species associated with inflammation were altered during extended room temperature storage. Interestingly, alterations were observed in apoptosis-associated ceramide species suggesting that cold storage of platelets may delay the progression of apoptosis. The lipidome of the cryopreserved platelets was not considerably altered immediately after thawing. However, changes became apparent during post-thaw storage. In contrast, the lipid profile of microparticles formed after thawing was significantly different to the lipid profile of the microparticles present prior to freezing. More specifically, externalisation of lipids associated with coagulation (PS and PE) were increased immediately after thawing. Further, lipid changes present in the post-thaw microparticles and the supernatant (LPC and LPE) may be associated with altered inflammation and signalling. Overall, the research presented in this dissertation has expanded the knowledge of alternatively stored platelet products and thus may be valuable in expanding their utility.
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