Platelet removal by single-step centrifugation

Rikkert, Linda Gerritdina; Coumans, F.A.W.; Hau, Chi M.; Terstappen, Leon W.M.M.; Nieuwland, Rienk

doi:10.1080/09537104.2020.1779924

Cited by 26 publications

(24 citation statements)

References 6 publications

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“…While care was taken in the preparation of plasma samples, so as to avoid the inclusion of platelets within the final sample, we did not include a second centrifugation step to eliminate any residual platelets from samples and as such this may have affected BDNF and other measures sensitive to platelet activity. However, preparation of plasma was carried out at higher speeds in a similar manner to a recently validated protocol for the removal of platelets by singlestep centrifugation (Rikkert et al, 2020), although this protocol utilized a 20-minute centrifugation, compared to the 10-minute period used within the current study.…”

Section: Limitations and Future Directionsmentioning

confidence: 99%

Cerebral Hemodynamic and Neurotrophic Factor Responses Are Dependent on the Type of Exercise

et al. 2021

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This study examined acute cerebral hemodynamic and circulating neurotrophic factor responses to moderate intensity continuous exercise (MICT), guideline-based high intensity interval exercise (HIIT), and sprint interval exercise (SIT). We hypothesized that the pattern of middle cerebral artery velocity (MCAv) response would differ between interval and continuous exercise, with SIT inducing the smallest increase from rest, while increases in neurotrophic factors would be intensity-dependent. In a randomized crossover design, 24 healthy adults (nine females) performed three exercise protocols: (i) MICT (30 min), (ii) HIIT (4 × 4 min at 85% HRmax), and (iii) SIT (4 × 30 s supramaximal). MCAv significantly increased from rest across MICT (Δ13.1 ± 8.5 cm⋅s–1, p < 0.001) and all bouts of HIIT (Δ15.2 ± 9.8 cm⋅s–1, p < 0.001), but only for the initial bout of SIT (Δ17.3 ± 11.6 cm⋅s–1, p < 0.001). Immediately following each interval bout, MCAv increased (i.e., rebounded) for the SIT (9–14% above rest, p ≤ 0.04), but not HIIT protocol. SIT alone induced significant elevations from rest to end-exercise in vascular endothelial growth factor (VEGF; Δ28 ± 36%, p = 0.017) and brain-derived neurotrophic factor (BDNF, Δ149% ± 162%, p < 0.001) and there were greater increases in lactate than in either other protocol (>5-fold greater in SIT, p < 0.001), alongside a small significant reduction at the end of active recovery in insulin-like growth factor 1 (IGF-1, Δ22 ± 21%, p = 0.002). In conclusion, while the nature of the response may differ, both guideline-based and sprint-based interval exercise have the potential to induce significant changes in factors linked to improved cerebrovascular and brain health.

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Section: Limitations and Future Directionsmentioning

confidence: 99%

Cerebral Hemodynamic and Neurotrophic Factor Responses Are Dependent on the Type of Exercise

et al. 2021

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“…Concerning the latter, platelets need special attention when studying blood EVs because they can be easily activated under blood collection, handling and storage, and release EVs that may confound the results [ 99 , 102 – 105 ]. Two subsequent centrifugations at 2500 × g for 15 min have often been used to deplete platelets from plasma samples, but a protocol using a single-step centrifugation has recently been proposed [ 98 , 102 , 106 ]. Moreover, blood samples contain lipoproteins of similar sizes to EVs [ 35 , 107 ].…”

Section: Evs As Liquid Biomarkers For Prostate Cancer: Methodological...mentioning

confidence: 99%

Extracellular vesicles as a source of prostate cancer biomarkers in liquid biopsies: a decade of research

et al. 2021

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Prostate cancer is a global cancer burden and considerable effort has been made through the years to identify biomarkers for the disease. Approximately a decade ago, the potential of analysing extracellular vesicles in liquid biopsies started to be envisaged. This was the beginning of a new exciting area of research investigating the rich molecular treasure found in extracellular vesicles to identify biomarkers for a variety of diseases. Vesicles released from prostate cancer cells and cells of the tumour microenvironment carry molecular information about the disease that can be analysed in several biological fluids. Numerous studies document the interest of researchers in this field of research. However, methodological issues such as the isolation of vesicles have been challenging. Remarkably, novel technologies, including those based on nanotechnology, show promise for the further development and clinical use of extracellular vesicles as liquid biomarkers. Development of biomarkers is a long and complicated process, and there are still not many biomarkers based on extracellular vesicles in clinical use. However, the knowledge acquired during the last decade constitutes a solid basis for the future development of liquid biopsy tests for prostate cancer. These are urgently needed to bring prostate cancer treatment to the next level in precision medicine.

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“…The phenomena taking place during centrifugation of blood are complex [41,42]. The prediction of particle concentration change with space and time in the final preparation is further complicated by the lack of precision in separation of PPP from PVRP (relevant when cells are not completely sedimented), by retention of platelets and EVs on the walls of the tube, and floatation of platelets in lipemic samples [43].…”

Section: Limitations Of the Studymentioning

confidence: 99%

Enrichment of plasma in platelets and extracellular vesicles by the counterflow to erythrocyte settling

et al. 2021

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In order to prepare optimal platelet and extracellular vesicle (EV)-rich plasma for the treatment of chronic temporal bone inflammation, we studied effects of centrifugation parameters on redistribution of blood constituents in blood samples of 23 patients and 20 volunteers with no record of disease. Concentrations of blood cells and EVs were measured by flow cytometry. Sample content was inspected by scanning electron microscopy. A mathematical model was constructed to interpret the experimental results. The observed enrichment of plasma in platelets and EVs after a single spin of blood depended on the erythrocyte sedimentation rate, thereby indicating the presence of a flow of plasma that carried platelets and EVs in the direction opposite to settling of erythrocytes. Prolonged handling time correlated with the decrease of concentration of platelets and larger EVs in platelet and EV-rich plasma (PVRP), R = −0.538, p = 0.003, indicating cell fragmentation during the processing of samples. In further centrifugation of the obtained plasma, platelet and EV enrichment depended on the average distance of the sample from the centrifuge rotor axis. Based on the agreement of the model predictions with observations, we propose the centrifugation protocol optimal for platelet and EV enrichment and recovery in an individual sample, adjusted to the dimensions of the centrifuge rotor, volume of blood and erythrocyte sedimentation rate.

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Platelet removal by single-step centrifugation

Cited by 26 publications

References 6 publications

Cerebral Hemodynamic and Neurotrophic Factor Responses Are Dependent on the Type of Exercise

Cerebral Hemodynamic and Neurotrophic Factor Responses Are Dependent on the Type of Exercise

Extracellular vesicles as a source of prostate cancer biomarkers in liquid biopsies: a decade of research

Enrichment of plasma in platelets and extracellular vesicles by the counterflow to erythrocyte settling

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