期刊
PANCREAS
卷 46, 期 2, 页码 252-259出版社
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1097/MPA.0000000000000717
关键词
pancreas; human; RNA quality; organ donors; diabetes; RIN
资金
- Network for Pancreatic Organ Donors with Diabetes
- Juvenile Diabetes Research Foundation [25-2013-268, 25-2012-380, 25-2007-874, 47-2013-520]
- National Institute of Diabetes Digestive and Kidney Diseases [DK104155-01]
Objectives: Attaining high-quality RNA fromthe 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 (Delta RIN = 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.
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