Siewiera 2021 Int J Mol Sci

From Bioblast
Publications in the MiPMap
Siewiera K, Labieniec-Watala M, Wolska N, Kassassir H, Watala C (2021) Sample preparation as a critical aspect of blood platelet mitochondrial respiration measurements - The impact of platelet activation on mitochondrial respiration.

Β» Int J Mol Sci 22:9332. PMID: 34502240 Open Access

Siewiera Karolina, Labieniec-Watala Magdalena, Wolska Nina, Kassassir Hassan, Watala Cezary (2021) Int J Mol Sci

Abstract: Blood platelets are considered as promising candidates as easily-accessible biomarkers of mitochondrial functioning. However, their high sensitivity to various stimulus types may potentially affect mitochondrial respiration and lead to artefactual outcomes. Therefore, it is crucial to identify the factors associated with platelet preparation that may lead to changes in mitochondrial respiration. A combination of flow cytometry and advanced respirometry was used to examine the effect of blood anticoagulants, the media used to suspend isolated platelets, respiration buffers, storage time and ADP stimulation on platelet activation and platelet mitochondria respiration. Our results clearly show that all the mentioned factors can affect platelet mitochondrial respiration. Briefly, (i) the use of EDTA as anticoagulant led to a significant increase in the dissipative component of respiration (LEAK), (ii) the use of plasma for the suspension of isolated platelets with MiR05 as a respiration buffer allows high electron transfer capacity and low platelet activation, and (iii) ADP stimulation increases physiological coupling respiration (ROUTINE). Significant associations were observed between platelet activation markers and mitochondrial respiration at different preparation steps; however, the fact that these relationships were not always apparent suggests that the method of platelet preparation may have a greater impact on mitochondrial respiration than the platelet activation itself. β€’ Keywords: Blood platelets, Mitochondria, Mitochondrial respiration, Platelet activation, Sample preparation β€’ Bioblast editor: Plangger M β€’ O2k-Network Lab: PL Lodz Watala C

Labels: MiParea: Respiration, Instruments;methods 

Organism: Human  Tissue;cell: Blood cells  Preparation: Intact cells 

Coupling state: LEAK, ROUTINE, ET  Pathway: ROX  HRR: Oxygraph-2k 


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