[Frontiers in Bioscience E3, 1162-1168, June 1, 2011]

Gene expression profiles of in vivo derived mouse blastocysts after slow-freezing

Li Wang1, Xinyan Zhang2, Chunhua Yu3, Yuanqing Yao2

1Peking University Stem Cell Research Center, Peking University, Beijing. P. R. China, 2Department of Obstetrics and Gynecology, the General Hospital of the People's Liberation Army, Beijing. P. R. China, 3Department of Gynecology and Obstetrics, Tangdu Hospital, The Fourth Military Medical University, Xi'an, PR China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Blastocysts collection
3.2. Slow freezing and thawing
3.3. RNA extraction and RNA Amplification
3.4. Hybridization and Slide Processing
3.5. Scanning and result analyses
3.6. Quantitative real-time RT-RCR
3.7. Statistical analysis
4. Results
4.1. The differential expression profile of cryopreserved blastocysts detected by microarray
4.2. Gene ontologies (GO) classification of differential genes
4.3. Pathway analysis of differential genes
4.4. Confirmation with qRT-PCR
5. Discussion
6. Acknowledgements
7. References

1. ABSTRACT

The goals of our study were to analyze change of global gene expression profile of mouse blastocysts after slow-freezing and to explore molecular mechanism underlying the decreased pregnancy rate caused by cryopreservation. The results showed that superovulation differentially regulated the expression of 288 genes with at least 2.0-fold change considerations. Among which, 275 genes were down-regulated and the remainder up-regulated in the cryopreserved group. The independent analysis with real-time PCR fully confirmed the results of microarray. These differential genes were classified into eighteen functional groups belonging to biological process, molecular function, and cellular component. Twenty nine genes could be categorized into one or more of KEGG pathways. The pathways of mitogen-activated protein kinase, Wnt and cell cycle were the most predominantly affected. Thus, the expression pattern reflected a broad spectrum of consequences of slow-freezing on the blastocysts, with most effects on stress-related and cell cycle-related genes.