Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Effects of Cold Ischemia on RNA Stability and Quality of Lung Tissues Based on Standard PREanalytical Code Categorization.

Author(s): Matsubara T, Tomida S, Soh J, Uwabo T, Mori Y, Morita M, Nasu Y, Kanazawa S, Toyooka S

Publication: Biopreserv Biobank, 2017, Vol. 15, Page 484-486

PubMed ID: 28910147 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study investigated the effects of cold ischemia time on RNA yield and integrity and explored potential correlations between Standard PREanalytical Code (SPREC)-defined categories of cold ischemia time and RNA quality indicators. Forty nonmalignant lung specimens distal to the tumor site were obtained during surgery and transported to the biobank at room temperature (time range: 8 - 108 min). Specimens were divided and fixed in RNAlater (processing time range 13 – 63 min). Cold ischemia time (25-123 min) consisted of transport and processing time for each specimen. Specimens were incubated overnight at 4°C and then stored at -80°C until RNA was extracted according to manufacturer’s instructions using an RNeasy Mini Kit. RNA yields were determined by spectrophotometry. RNA integrity was assessed by GAPDH expression analysis by real-time qRT-PCR and by RIN calculation using High Sensitivity RNA ScreenTape of electropherograms obtained from an Agilent 2200 TapeStation. Statistical analysis was performed to determine the correlations between RNA quality indicators and SPREC-defined categories of cold ischemia time (20–30 min, n=3; 30–60 min, n=29); >60 min, n=8).

   Summary of Findings:

   Cold ischemia times ranging from 25 to 123 minutes was not statistically correlated with the RNA integrity indicators of RIN or GAPDH qRT-PCR amplification and no significant correlations were observed between these indicators and transport time (range: 8 – 108 min) or processing time (range: 13 – 63 min). Statistical analysis revealed no correlation between RNA integrity indicators and long or short transport times (not defined) and long or short processing times (not defined). In addition, there was no statistical correlation between RNA integrity indicators and SPREC-defined categories of cold ischemia.

   Biospecimens

   • Tissue - Lung

   Preservative Types

   • RNAlater

   Diagnoses:

   • Neoplastic - Normal Adjacent

   Platform:

   Analyte	Technology Platform
   RNA	Spectrophotometry
   RNA	Electrophoresis
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Acquisition	Cold ischemia time	25-123 min
   Storage	Specimen transport duration/condition	8 – 108 min
   Biospecimen Preservation	Duration of tissue/ specimen processing	13 – 63 min
   Real-time qRT-PCR Specific	Targeted nucleic acid	GAPDH

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