Literature DB >> 24951174

Banking placental tissue: an optimized collection procedure for genome-wide analysis of nucleic acids.

L M Wolfe1, R D Thiagarajan1, F Boscolo2, V Taché3, R L Coleman4, J Kim5, W K Kwan1, J F Loring4, M Parast5, L C Laurent6.   

Abstract

INTRODUCTION: Banking of high-quality placental tissue specimens will enable biomarker discovery and molecular studies on diseases involving placental dysfunction. Systematic studies aimed at developing feasible standardized methodology for placental collection in a typical clinical setting are lacking.
METHODS: To determine the acceptable timeframe for placental collection, we collected multiple samples from first and third trimester placentas at serial timepoints in a 2-h window after delivery, simultaneously comparing the traditional snap-freeze technique to commercial solutions designed to preserve RNA (RNAlater™), and DNA (DNAgard(®)). The performance of RNAlater for preserving DNA was also tested. Nucleic acid quality was assessed by determining the RNA integrity number (RIN) and genome-wide microarray profiling for gene expression and DNA methylation.
RESULTS: We found that samples collected in RNAlater had higher and more consistent RINs compared to snap-frozen tissue. Similar RINs were obtained for tissue collected in RNAlater as large (1 cm(3)) and small (∼0.1 cm(3)) pieces. RNAlater appeared to better stabilize the time zero gene expression profile compared to snap-freezing for first trimester placenta. DNA methylation profiles remained quite stable over a 2 h time period after removal of the placenta from the uterus, with DNAgard being superior to other treatments. DISCUSSION AND
CONCLUSION: The collection of placental samples in RNAlater and DNAgard is simple, and eliminates the need for liquid nitrogen or a freezer on-site. Moreover, the quality of the nucleic acids and the resulting data from samples collected in these preservation solutions is higher than samples collected using the snap-freeze method and easier to implement in busy clinical environments.
Copyright © 2014 Elsevier Ltd. All rights reserved.

Entities:  

Keywords:  DNA methylation; DNAgard; Gene expression; Microarray; Placenta; RNA integrity; RNAlater; Snap-freezing

Mesh:

Substances:

Year:  2014        PMID: 24951174      PMCID: PMC4847722          DOI: 10.1016/j.placenta.2014.05.005

Source DB:  PubMed          Journal:  Placenta        ISSN: 0143-4004            Impact factor:   3.481


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