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Pancreas. 2017 Feb;46(2):252-259. doi: 10.1097/MPA.0000000000000717.

Factors That Influence the Quality of RNA From the Pancreas of Organ Donors.

Author information

1
From the *Departments of Pathology, Immunology, & Laboratory Medicine, University of Florida, Gainesville, FL; †Department of Medicine, College of Medicine, University of Tennessee Health Science Center, Memphis, TN; ‡Diabetes Research Institute and Departments of Medicine, Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, FL; §Department of Information Sciences, ∥Division of Biostatistics, City of Hope, Duarte, CA; and ¶Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL.

Abstract

OBJECTIVES:

Attaining high-quality RNA from the tissues or organs of deceased donors used for research can be challenging due to physiological and logistical considerations. In this investigation, METHODS: RNA Integrity Number (RIN) was determined in pancreatic samples from 236 organ donors and used to define high (≥6.5) and low (≤4.5) quality RNAs. Logistic regression was used to evaluate the potential effects of novel or established organ and donor factors on RIN.

RESULTS:

Univariate analysis revealed donor cause of death (odds ratio [OR], 0.35; 95% confidence interval [CI], 0.15-0.77; P = 0.01), prolonged tissue storage before RNA extraction (OR, 0.65; 95% CI, 0.52-0.79; P < 0.01), pancreas region sampled (multiple comparisons, P < 0.01), and sample type (OR, 0.32; 95% CI, 0.15-0.67; P < 0.01) negatively influenced outcome. Conversely, duration of final hospitalization (OR, 3.95; 95% CI, 1.59-10.37; P < 0.01) and sample collection protocol (OR, 8.48; 95% CI, 3.96-19.30; P < 0.01) positively impacted outcome. Islet RNA obtained via laser capture microdissection improved RIN when compared with total pancreatic RNA from the same donor (ΔRIN = 1.3; 95% CI, 0.6-2.0; P < 0.01).

CONCLUSIONS:

A multivariable model demonstrates that autopsy-free and biopsy-free human pancreata received, processed, and preserved at a single center, using optimized procedures, from organ donors dying of anoxia with normal lipase levels increase the odds of obtaining high-quality RNA.

PMID:
27984510
PMCID:
PMC5235995
DOI:
10.1097/MPA.0000000000000717
[Indexed for MEDLINE]
Free PMC Article

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