Dataset Information

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

ABSTRACT: 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.

SUBMITTER: Eswaran N

PROVIDER: S-EPMC5815974 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

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

Eswaran Nalini N Agaram Sundaram Vickram V Rao Kamini Arvind KA Thalaivarisai Balasundaram Sridharan S

3 Biotech 20180216 3

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 pres ...[more]

PMID: 29484278

Similar Datasets

Project description:BackgroundExtracellular vesicles (EVs) are essential for cell-to-cell communication because they transport functionally active molecules, including proteins, RNA, and lipids, from secretory cells to nearby or distant target cells. Seminal plasma contains a large number of EVs (sEVs) that are phenotypically heterogeneous. The aim of the present study was to identify the RNA species contained in two subsets of porcine sEVs of different sizes, namely small sEVs (S-sEVs) and large sEVs (L-sEVs). The two subsets of sEVs were isolated from 54 seminal plasma samples by a method combining serial centrifugations, size exclusion chromatography, and ultrafiltration. The sEVs were characterized using an orthogonal approach. Analysis of RNA content and quantification were performed using RNA-seq analysis.ResultsThe two subsets of sEVs had different size distributions (P < 0.001). They also showed differences in concentration, morphology, and specific protein markers (P < 0.05). A total of 735 RNAs were identified and quantified, which included: (1) mRNAs, rRNAs, snoRNAs, snRNAs, tRNAs, other ncRNAs (termed as "all RNAs"), (2) miRNAs and (3) piRNAs. The distribution pattern of these RNA classes differed between S-sEVs and L-sEVs (P < 0.05). More than half of "all RNAs", miRNAs and piRNAs were found to be differentially abundant between S- and L-sEVs (FDR < 0.1%). Among the differentially abundant RNAs, "all RNAs" were more abundant in L- than in S-sEVs, whereas the most of the miRNAs were more abundant in S- than in L-sEVs. Differentially abundant piRNAs were equally distributed between S- and L-sEVs. Some of the all RNAs and miRNAs found to be differentially abundant between S- and L-sEVs were associated with sperm quality and functionality and male fertility success.ConclusionsSmall and large sEVs isolated from porcine seminal plasma show quantitative differences in RNA content. These differences would suggest that each sEV subtype exerts different functional activities in the targeted cells, namely spermatozoa and functional cells of the female reproductive tract.

Dataset Information

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

Publications

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

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets