Simple isolation and characterization of seminal plasma extracellular vesicle and its total RNA in an academic lab

Extracellular vesicles (EVs) are small membrane-bound sacs, identified in many body fluids of humans. Standard extracellular vesicle separation methods such as differential and ultracentrifugation are very expensive, not affordable in academic labs. So, the current research tried to isolate seminal...

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Bibliographic Details
Published in:3 Biotech 2018-03, Vol.8 (3), p.139-6, Article 139
Main Authors: Eswaran, Nalini, Agaram Sundaram, Vickram, Rao, Kamini Arvind, Thalaivarisai Balasundaram, Sridharan
Format: Article
Language:English
Subjects:
Agriculture
Bioinformatics
Biomaterials
Biotechnology
Body fluids
Cancer Research
Chemistry
Chemistry and Materials Science
Fertility
Membrane vesicles
Polyethylene glycol
Ribonucleic acid
RNA
Semen
Short Reports
Stem Cells
Ultracentrifugation
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Summary:Extracellular vesicles (EVs) are small membrane-bound sacs, identified in many body fluids of humans. Standard extracellular vesicle separation methods such as differential and ultracentrifugation are very expensive, not affordable in academic labs. So, the current research tried to isolate seminal plasma EVs using polyethylene glycol (PEG) precipitation process. Normospermia semen from “Milann - The Fertility Center” processed to isolate EVs by PEG method. Nanodrop spectrophotometer showed presence of EVs by indirectly measuring protein content of precipitated EVs. EVs isolated by PEG precipitation showed a wide size range from 30 to 1000 nm with Z average of 75.4 nm and a PI of 0.464, whereas ultracentrifuge sample showed size range of 60–1000 nm with Z average of 501.3 nm with a PI of 0.692. Edax analysis also showed good elemental pattern. Total RNA extraction from PEG EVs analysed with nanodrop spectrophotometer, showed presence of RNA content in varying concentrations obtained from different ratios in nanograms. Thus, the current study concludes that seminal plasma EVs isolated by PEG precipitation is simple, reproducible and non-sensitive to carry out at academic labs.
ISSN:2190-572X
2190-5738
DOI:10.1007/s13205-018-1157-7