Abstract
Collecting biological tissue samples in a biobank grants a unique opportunity to validate diagnostic and therapeutic strategies for translational and clinical research. 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, thus far, 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 (January 2004–December 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. FFPE biospecimens demonstrated unaltered RNA stability over time for up to 10 years. Furthermore, we provide a protocol for processing tissue samples in our Munich Vascular Biobank. In this work, we demonstrate that biobanking is an important tool not only for scientific research but also for clinical usage and personalized medicine.
Highlights
Biobanking is considered as a tool to collect and store biological tissue samples, mainly for research purposes
* Starting from 2016, we started to focus on peripheral aneurysm and thrombus as well (PAD/aneurysm/thrombus). ** Since 2017, we have focused only on peripheral aneurysm and thrombus. # abdominal aortic aneurysm (AAA) tissue is increasingly difficult to obtain due to the fact that open aneurysm repair is more and more frequently replaced by endovascular techniques
Our results demonstrated no significant differences in RNA integrity number (RIN) for up to 10 years back, indicating that the RNA is stable over time even if the fixed paraffin-embedded specimens (FFPE) biospecimens are stored at room temperature
Summary
Biobanking is considered as a tool to collect and store biological tissue samples, mainly for research purposes. New biobanks have been built focusing on selective human tissues, such as various cancer types or atherosclerotic specimens [1,2,3] In this context, the collection of human atherosclerotic biomaterial following, for example, surgical intervention of stenotic carotid arteries or excision of aortic wall from patients with abdominal aortic aneurysm (AAA), has become a important issue to improve our understanding of the underlying mechanisms leading to the formation and progression of atherosclerotic plaques and aneurysm toward vulnerable and rupture-prone phenotype. Renu Virmani et al investigated coronary arteries from patients with sudden cardiac death and characterized additional plaque features, such as erosion, calcified nodules, fibrous cap atheroma, and fibrocalcific plaques [11] All these studies were based on observations of large atherosclerotic tissue cohorts collected over years in the corresponding biobanks [12]. We provide our long-standing experience in running a biobank of vascular tissue samples, including our analyses of tissue quality from FFPE samples, for successful and reliable expression analyses
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