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Evaluation of platelet mitochondria integrity after treatment with Mirasol pathogen reduction technology
Junzhi Li1, Owen Lockerbie, Dirk de Korte
1Navigant Biotechnologies, Inc., Lakewood, Colorado 80215, USA.
Background:
Previous studies showed that Mirasol (Navigant Biotechnologies, Inc.) pathogen reduction technology (PRT) treatment resulted in an increase in platelet (PLT) glucose consumption and lactate production rates and decrease in pH in media during PLT storage. Increased glycolytic flux could result from damage to mitochondria and/or increased ATP consumption.
Study Design And Methods:
PLT concentrates were collected by standard automated blood component collection system (Trima, Gambro BCT) procedure on Day 0 and treated with Mirasol PRT treatment on Day 1. PLT mitochondrial transmembrane potential was evaluated by staining PLTs with JC-1 followed by flow cytometry analysis. Mitochondrial enzymatic activity was measured by the MTT assay. ATP content and pH were also quantified. The values for these measurements were compared among control, untreated, and pathogen reduction technology (PRT)-treated PLTs during PLT storage for up to 7 days.
Results:
No significant changes were found in pH, JC-1 signal, MTT activity, and ATP content of the PLTs immediately after PRT treatment. The treated PLTs exhibited a moderate but significantly accelerated decrease in pH and lower ATP content after 7-day storage when compared to control PLTs. Neither the JC-1 assay nor the MTT assay, however, showed a significant difference between control and treated PLTs during PLT storage.
Conclusions:
There is no evidence from these studies that Mirasol PRT treatment alters PLT mitochondrial structural and functional integrity immediately after treatment and during PLT storage. An increased demand for ATP may be the driving force for observed increases in both the glycolytic flux and the oxidative metabolism observed in treated PLTs.
Insights
Mirasol pathogen reduction technology (PRT) does not harm platelet mitochondria. Treated platelets show increased metabolic activity and lower ATP during storage, suggesting a higher ATP demand.
Area of Science:
- Blood component processing
- Transfusion medicine
- Biotechnology
Background:
- Previous studies indicated Mirasol pathogen reduction technology (PRT) increases platelet (PLT) glucose consumption and lactate production, decreasing media pH.
- This suggests increased glycolytic flux due to mitochondrial damage or higher ATP consumption.
Purpose of the Study:
- To evaluate the impact of Mirasol PRT on platelet mitochondrial integrity and function.
- To assess changes in platelet mitochondrial transmembrane potential, enzymatic activity, ATP content, and pH during storage post-treatment.
Main Methods:
- Platelet concentrates were collected and treated with Mirasol PRT.
- Mitochondrial transmembrane potential assessed via JC-1 staining and flow cytometry.
- Mitochondrial enzymatic activity measured by MTT assay.
- ATP content and pH quantified over 7 days of storage.
Main Results:
- No immediate changes in platelet pH, JC-1 signal, MTT activity, or ATP content post-PRT treatment.
- After 7-day storage, PRT-treated platelets showed significantly lower ATP content and accelerated pH decrease compared to controls.
- JC-1 and MTT assays revealed no significant differences in mitochondrial integrity or activity between control and treated platelets during storage.
Conclusions:
- Mirasol PRT treatment does not appear to alter platelet mitochondrial structural or functional integrity.
- Observed increases in glycolytic flux and oxidative metabolism in treated platelets may be driven by an increased demand for ATP.
