Swiss Vascular Biobank: Evaluation of Optimal Extraction Method and Admission Solution for Preserving RNA from  Human Vascular Tissue

Author:

Pelisek Jaroslav1ORCID,Yundung Yankey1ORCID,Reutersberg Benedikt1ORCID,Meuli Lorenz1ORCID,Rössler Fabian2ORCID,Rabin Laetitia1,Kopp Reinhard1,Zimmermann Alexander1ORCID

Affiliation:

1. Department of Vascular Surgery, University Hospital Zurich, 8091 Zurich, Switzerland

2. Department of Surgery and Transplantation, University Hospital Zurich, 8091 Zurich, Switzerland

Abstract

Proper biobanking is essential for obtaining reliable data, particularly for next-generation sequencing approaches. Diseased vascular tissues, having extended atherosclerotic pathologies, represent a particular challenge due to low RNA quality. In order to address this issue, we isolated RNA from vascular samples collected in our Swiss Vascular Biobank (SVB); these included abdominal aortic aneurysm (AAA), peripheral arterial disease (PAD), healthy aorta (HA), and muscle samples. We used different methods, investigated various admission solutions, determined RNA integrity numbers (RINs), and performed expression analyses of housekeeping genes (ACTB, GAPDH), ribosomal genes (18S, 28S), and long non-coding RNAs (MALAT1, H19). Our results show that RINs from diseased vascular tissue are low (2–4). If the isolation of primary cells is intended, as in our SVB, a cryoprotective solution is a better option for tissue preservation than RNAlater. Because RNA degradation proceeds randomly, controls with similar RINs are recommended. Otherwise, the data might convey differences in RNA degradation rather than the expressions of the corresponding genes. Moreover, since the 18S and 28S genes in the diseased vascular samples were degraded and corresponded with the low RINs, we believe that DV200, which represents the total RNA’s disintegration state, is a better decision-making aid in choosing samples for omics analyses.

Funder

Department of Vascular Surgery

Publisher

MDPI AG

Subject

General Medicine

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