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 language | English (US) |
|---|---|
| Pages (from-to) | 252-259 |
| Number of pages | 8 |
| Journal | Pancreas |
| Volume | 46 |
| Issue number | 2 |
| DOIs | |
| State | Published - 2017 |
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Keywords
- Diabetes
- Human
- Organ donors
- Pancreas
- RIN
- RNA quality
ASJC Scopus subject areas
- Internal Medicine
- Endocrinology, Diabetes and Metabolism
- Hepatology
- Endocrinology
Cite this
Factors that influence the quality of RNA from the pancreas of organ donors. / Philips, Tiffany; Kusmartseva, Irina; Gerling, Ivan C.; Campbell-Thompson, Martha; Wasserfall, Clive; Pugliese, Alberto; Longmate, Jeffrey A.; Schatz, Desmond A.; Atkinson, Mark A.; Kaddis, John S.
In: Pancreas, Vol. 46, No. 2, 2017, p. 252-259.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Factors that influence the quality of RNA from the pancreas of organ donors
AU - Philips, Tiffany
AU - Kusmartseva, Irina
AU - Gerling, Ivan C.
AU - Campbell-Thompson, Martha
AU - Wasserfall, Clive
AU - Pugliese, Alberto
AU - Longmate, Jeffrey A.
AU - Schatz, Desmond A.
AU - Atkinson, Mark A.
AU - Kaddis, John S.
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Diabetes
KW - Human
KW - Organ donors
KW - Pancreas
KW - RIN
KW - RNA quality
UR - http://www.scopus.com/inward/record.url?scp=85006263045&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85006263045&partnerID=8YFLogxK
U2 - 10.1097/MPA.0000000000000717
DO - 10.1097/MPA.0000000000000717
M3 - Article
C2 - 27984510
AN - SCOPUS:85006263045
VL - 46
SP - 252
EP - 259
JO - Pancreas
JF - Pancreas
SN - 0885-3177
IS - 2
ER -