Back to Search Start Over

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

Authors :
Wolfe, L M
Thiagarajan, R D
Boscolo, F
Taché, V
Coleman, R L
Kim, J
Kwan, W K
Loring, J F
Parast, M
Laurent, L C
Source :
Placenta; 2014 Aug, Vol. 35 Issue 8, p645-654, 10p
Publication Year :
2014

Abstract

<bold>Introduction: </bold>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.<bold>Methods: </bold>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.<bold>Results: </bold>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.<bold>Discussion and Conclusion: </bold>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. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
01434004
Volume :
35
Issue :
8
Database :
Supplemental Index
Journal :
Placenta
Publication Type :
Academic Journal
Accession number :
107798197
Full Text :
https://doi.org/10.1016/j.placenta.2014.05.005