Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program
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ORIGINAL PAPER
Characterization of RNA isolated from eighteen different human tissues: results from a rapid human autopsy program Douglas G. Walker . Alexis M. Whetzel . Geidy Serrano . Lucia I. Sue . Lih-Fen Lue . Thomas G. Beach
Received: 7 May 2015 / Accepted: 7 April 2016 Ó Springer Science+Business Media Dordrecht 2016
Abstract Many factors affect the integrity of messenger RNA from human autopsy tissues including postmortem interval (PMI) between death and tissue preservation and the pre-mortem agonal and disease states. In this communication, we describe RNA isolation and characterization of 389 samples from 18 different tissues from elderly donors who were participants in a rapid whole-body autopsy program located in Sun City, Arizona (www.brainandbodydona tionprogram.org). Most tissues were collected within a PMI of 2–6 h (median 3.15 h; N = 455), but for this study, tissue from cases with longer PMIs (1.25–29.25 h) were included. RNA quality was assessed by RNA integrity number (RIN) and total yield (ng RNA/mg tissue). RIN correlated with PMI for heart (r = -0.531, p = 0.009) and liver (r = -558, p = 0.0017), while RNA yield correlated with PMI for colon (r = -485, p = 0.016) and skin (r = -0.460, p = 0.031). RNAs with the lowest integrity were from skin and cervix where 22.7 and
31.4 % of samples respectively failed to produce intact RNA; by contrast all samples from esophagus, lymph node, jejunum, lung, stomach, submandibular gland and kidney produced RNA with measurable RINs. Expression levels in heart RNA of 4 common housekeeping normalization genes showed significant correlations of Ct values with RIN, but only one gene, glyceraldehyde-3 phosphate dehydrogenase, showed a correlation of Ct with PMI. There were no correlations between RIN values obtained for liver, adrenal, cervix, esophagus and lymph node and those obtained from corresponding brain samples. We show that high quality RNA can be produced from most human autopsy tissues, though with significant differences between tissues and donors. The RNA stability and yield did not depend solely on PMI; other undetermined factors are involved, but these do not include the age of the donor. Keywords RNA integrity Aging Gene expression Postmortem interval Autopsy Human Tissue
D. G. Walker A. M. Whetzel G. Serrano L. I. Sue T. G. Beach Banner Sun Health Research Institute, Sun City, AZ, USA
Introduction
Present Address: D. G. Walker (&) L.-F. Lue Laboratory of Neuroinflammation, Biodesign Neurodegenerative Disease Research Center, Arizona State University, Room 538, School of Life Sciences – E Wing, 427 E. Tyler Mall, Tempe, AZ 85287, USA e-mail: [email protected]
Since the decoding of the human genome and development of methods to survey transcripts of the whole genome, there has been an increasing need for high quality RNA from human tissue samples. Global gene expression profiling by microarrays or sequencing of
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Cell Tissue Bank
RNA from normal and diseased human tissues has been a key approach for many ye
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