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Updated: Mar 21, 2026
Isolation of High-density Lipoproteins for Non-coding Small RNA Quantification
Published on: November 28, 2016
Stability of serum high-density lipoprotein-microRNAs for preanalytical conditions
Hiroaki Ishikawa1, Hiroya Yamada2, Nao Taromaru3
11 Department of Biochemistry, Fujita Health University School of Health Sciences, Toyoake, Japan.
Abstract:
Background Recently, several studies have shown that microRNAs are present in high-density lipoprotein, and high-density lipoprotein-microRNA may be a promising disease biomarker. We investigated the stability of high-density lipoprotein-microRNAs in different storage conditions as this is an important issue for its application to the field of clinical research. Methods microRNAs were extracted from the high-density lipoprotein fraction that was purified from the serum. miR-135 a and miR-223, which are known to be present in high-density lipoprotein, were quantified by quantitative real-time PCR. The influence of preanalytical parameters on the analysis of high-density lipoprotein-miRNAs was examined by the effect of RNase, storage conditions, and freezing and thawing. Results The concentrations of microRNA in high-density lipoprotein were not altered by RNase A treatment (0-100 U/mL). No significant change in these microRNAs was observed after storing serum at room temperature or 4℃ for 0-24 h, and there was a similar result in the cryopreservation for up to two weeks. Also, high-density lipoprotein-microRNAs were stable for, at least, up to five freeze-thaw cycles. Conclusions These results demonstrated that high-density lipoprotein-microRNAs are relatively resistant to various storage conditions. This study provides new and important information on the stability of high-density lipoprotein-microRNAs.
Insights
High-density lipoprotein-microRNAs show remarkable stability across various storage conditions, including RNase treatment, extended room temperature or refrigerated storage, and multiple freeze-thaw cycles. This resilience supports their potential as reliable disease biomarkers in clinical research.
Area of Science:
- Biochemistry
- Molecular Biology
- Clinical Diagnostics
Background:
- MicroRNAs (miRNAs) are increasingly recognized within high-density lipoprotein (HDL) particles.
- HDL-miRNAs are emerging as potential biomarkers for various diseases.
- Assessing the stability of HDL-miRNAs under preanalytical conditions is crucial for their clinical application.
Purpose of the Study:
- To evaluate the stability of high-density lipoprotein-microRNAs (HDL-miRNAs) under different storage and handling conditions.
- To determine the impact of RNase treatment, storage temperature, and freeze-thaw cycles on HDL-miRNA integrity.
Main Methods:
- MicroRNAs were isolated from the HDL fraction of human serum.
- Quantitative real-time PCR was used to measure the concentrations of miR-135a and miR-223.
- Stability was assessed following RNase A treatment, storage at room temperature and 4°C, cryopreservation, and multiple freeze-thaw cycles.
Main Results:
- HDL-miRNA concentrations remained stable after RNase A treatment (0-100 U/mL).
- No significant changes were observed after storing serum at room temperature or 4°C for up to 24 hours.
- HDL-miRNAs were stable during cryopreservation for two weeks and up to five freeze-thaw cycles.
Conclusions:
- High-density lipoprotein-microRNAs exhibit significant resistance to various preanalytical storage conditions.
- These findings provide critical data supporting the use of HDL-miRNAs as stable and reliable biomarkers in clinical research.
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