Factors that influence the quality of RNA from the pancreas of organ donors

Tiffany Philips, Irina Kusmartseva, Ivan C. Gerling, Martha Campbell-Thompson, Clive Wasserfall, Alberto Pugliese, Jeffrey A. Longmate, Desmond A. Schatz, Mark A. Atkinson, John S. Kaddis

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

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 (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.

Original languageEnglish (US)
Pages (from-to)252-259
Number of pages8
JournalPancreas
Volume46
Issue number2
DOIs
StatePublished - 2017

Keywords

  • Diabetes
  • Human
  • Organ donors
  • Pancreas
  • RIN
  • RNA quality

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

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  • Cite this

    Philips, T., Kusmartseva, I., Gerling, I. C., Campbell-Thompson, M., Wasserfall, C., Pugliese, A., Longmate, J. A., Schatz, D. A., Atkinson, M. A., & Kaddis, J. S. (2017). Factors that influence the quality of RNA from the pancreas of organ donors. Pancreas, 46(2), 252-259. https://doi.org/10.1097/MPA.0000000000000717