title: Factors That Influence the Quality of RNA From the Pancreas of Organ Donors.
abstract: 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.
  
  
  
  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).
  
  
  
  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.
authors.affiliation: 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.
authors.forename: Tiffany
authors.initials: T
authors.lastname: Philips
authors.forename: Irina
authors.initials: I
authors.lastname: Kusmartseva
authors.forename: Ivan C
authors.initials: IC
authors.lastname: Gerling
authors.forename: Martha
authors.initials: M
authors.lastname: Campbell-Thompson
authors.forename: Clive
authors.initials: C
authors.lastname: Wasserfall
authors.forename: Alberto
authors.initials: A
authors.lastname: Pugliese
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Longmate
authors.forename: Desmond A
authors.initials: DA
authors.lastname: Schatz
authors.forename: Mark A
authors.initials: MA
authors.lastname: Atkinson
authors.forename: John S
authors.initials: JS
authors.lastname: Kaddis
mesh_terms: D000293:Adolescent; D000328:Adult; D001344:Autopsy; D005260:Female; D006801:Humans; D016015:Logistic Models; D008297:Male; D008875:Middle Aged; D015999:Multivariate Analysis; D010179:Pancreas / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D020871:RNA Stability; D014019:Tissue Donors; D055815:Young Adult
chemical_list: D012313:RNA