Autor:innen:
Prof. Dr. Jaroslav Pelisek | Klinikum rechts der Isar der Technischen Universität München | Germany
Jessica Pauli | Klinikum rechts der Isar der Technischen Universität München | Germany
Sabine Bauer | Klinikum rechts der Isar der Technischen Universität München | Germany
Susanne Metschl | Klinikum rechts der Isar der Technischen Universität München | Germany
PD Dr. med. habil. Albert Busch | Klinikum rechts der Isar der Technischen Universität München | Germany
Dr. med. Benedikt Reutersberg | Klinikum rechts der Isar der Technischen Universität München | Germany
Dr. med. Michael Kallmayer | Klinikum rechts der Isar der Technischen Universität München | Germany
PD Dr. med. habil. Matthias Trenner | Klinikum rechts der Isar der Technischen Universität München | Germany
Dr. med. Heiko Wendorff | Klinikum rechts der Isar der Technischen Universität München | Germany
Dr. med. habil. Pavlos Tsantilas | Klinikum rechts der Isar der Technischen Universität München | Germany
Dr. med. Sofie Schmid | Klinikum rechts der Isar der Technischen Universität München | Germany
Prof. Dr. Lars Mädgefessel | Klinikum rechts der Isar der Technischen Universität München | Germany
Prof. Dr. med. Hans-Henning Eckstein | Klinikum rechts der Isar der Technischen Universität München | Germany
Collecting biological tissue samples in a biobank grants a unique opportunity to validate diagnostic and therapeutic strategies for translational and clinical research, including disease-related specific biomarkers, next-generation sequencing and omics analyses. In the present work, we provide our long-standing experience in establishing and maintaining a biobank of vascular tissue samples, including the evaluation of tissue quality, especially in formalin-fixed paraffin-embedded specimens (FFPE).
Our Munich Vascular Biobank includes vascular biomaterial from patients with high-grade carotid artery stenosis (n = 1567), peripheral arterial disease (n = 703), and abdominal aortic aneurysm (n = 481) from our Department of Vascular and Endovascular Surgery (2004–2018). Vascular tissue samples are continuously processed and characterized to assess tissue morphology, histological quality, cellular composition, inflammation, calcification, neovascularization, and the content of elastin and collagen fibers. Atherosclerotic plaques are further classified in accordance with the American Heart Association (AHA), and plaque stability is determined. In order to assess the quality of RNA from FFPE tissue samples over time (2009–2018), RNA integrity number (RIN) and the extent of RNA fragmentation were evaluated. Expression analysis was performed with two housekeeping genes—glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin (ACTB)—using TaqMan-based quantitative reverse-transcription polymerase chain reaction (qRT)-PCR.
All FFPE biospecimens demonstrated unaltered RIN independent of the sample age. The maximal length of RNA from the FFPE biospecimens tested in this study exceeded in most cases 500 nt, with many being even longer than 1000 nt. Furthermore, no significant differences in the overall RNA fragmentation were observed over time. Independent of the results from the Bioanalyzer (RIN, overall RNA fragmentation) expression of GAPDH and ACTB on mRNA level was detected in all biospecimens tested. The expression level depended only on the concentration of the total amount of extracted RNA and did not change significantly over the last 10 years.
In the present work, we summarize our own experiences in managing a vascular biobank, starting with tissue excision through different processing techniques, testing the quality of the biospecimens, up to their proper storage. Furthermore, evidence is provided that FFPE tissue samples are suitable not only for histological and immunohistochemical analyses but also for expression analyses at mRNA level and potential RNA sequencing. Biobanking of high-quality human biospecimens, together with patient clinical information, provides a fundamental scientific infrastructure for personalized medicine.