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In-line leukoreduction filters: A new source of microparticle for human and animal study
Mohammad Reza Javan1, Sedigheh Amini Kafi-Abad1, Mahin Nikougoftar Zarif2
1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Introduction:
The isolation of microparticles (MPs) from leukoreduction filters (LRFs) during cell extraction process introduced LRFs as a precious source of MPs for animal and human study.
Method:
LRFs were collected from Tehran Blood Transfusion Center. The back-flushing method was used for leukocyte extraction from the LRFs. MPs were isolated through double-step centrifugation. Dynamic light scattering (DLS), electron microscopy (EM), and flow cytometry were performed for the evaluation of MPs size, morphology, and structural properties respectively. Statistical analyses were carried out to evaluation of differences between test and control groups. a p-value less than 0.05 indicates significant differences.
Result:
DLS analysis showed that the average MP size in the test and control groups was 654.83 nm and 233.68 nm respectively. SEM images showed the spherical, oval, cell fragment, and micro-aggregate particles and TEM images demonstrated the mitochondrial-like body in the MPs. Flow cytometry studies also showed a significant increase in the percent of CD41, and CD14, and a significant decrease in the percent of CD235a in the test group compared to control (P value=0.029, P value=0.035, P value= 0.001 respectively). Moreover, the percentage of CD34 MPs indicated a borderline difference between the two groups (P value= 0.075). Finally count of MPs in the test and control groups was 1202095.34 and 280948.64, respectively and the difference was significant (P value=0.008).
Conclusion:
It is concluded that LRFs are a potential source of the large volume of various cell MPs with different phenotypical and structural properties for animal and human phase studies. Moreover, the investigation of LRFs as a source of different types of exosomes can shed new light on extracellular vesicle studies.
Insights
Leukoreduction filters (LRFs) are a valuable source of microparticles (MPs) for research. These MPs, isolated from LRFs, possess diverse characteristics suitable for various studies.
Area of Science:
- Biomedical research
- Extracellular vesicles
- Blood product analysis
Background:
- Leukoreduction filters (LRFs) are increasingly recognized as a source of microparticles (MPs).
- MPs isolated from LRFs offer potential for both animal and human studies.
Purpose of the Study:
- To evaluate LRFs as a source of microparticles (MPs).
- To characterize the size, morphology, and phenotype of MPs isolated from LRFs.
Main Methods:
- MPs were extracted from LRFs using a back-flushing method.
- Isolation involved double-step centrifugation, followed by characterization using Dynamic Light Scattering (DLS), Electron Microscopy (EM), and flow cytometry.
- Statistical analysis was performed to compare test and control groups.
Main Results:
- DLS revealed larger average MP size in the test group (654.83 nm) compared to the control (233.68 nm).
- Flow cytometry showed significant differences in CD41, CD14, and CD235a expression.
- A significantly higher MP count was observed in the test group (1,202,095.34) versus the control (280,948.64).
Conclusions:
- LRFs are a promising source for obtaining a large volume of diverse cell microparticles.
- The characterized MPs from LRFs possess distinct phenotypical and structural properties for preclinical and clinical studies.
- Further investigation into LRF-derived exosomes could advance extracellular vesicle research.
