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Differential Gene Expression Analysis of Pure Cell Populations from Frozen UterineTissue Using Laser Capture Microdissection (LCM) and cDNA Microarrays

Se-Jin Yoon and Kyung-Ah Lee Infertility Medical Center, CHA General Hospital, Seoul 135-081, Korea


Abstract
Laser Capture Microdissection (LCM) is an innovative technique which permits the rapid and reliable procurement of pure cell populations from heterogeneous tissue sections. After LCM, RNA isolated from pure cell populations was amplified to generate adequate amounts of antisense RNA (aRNA) for hybridization, and the differential gene expression profiles were analyzed between two experimental groups.


Introduction
When investigating molecular mechanisms in specific tissues or cells, the accuracy of deduced mechanisms often depends on the relative abundance of the cell population in question. Mammalian uterine tissue consists of endometrium and myometrium. The endometrium is comprised of luminal epithelium, densely packed stroma, and the germinal basalis. For implantation, blastocysts make their first physical uterine contact with the maternal endometrial luminal epithelium. In previous analyses of global gene expression during implantation, authors used whole uterine tissue, thus these studies produced information on the temporal gene expression profile in the uterus, but not in the specific cell population.1,2 Using LCM, a pure population of luminal epithelial tissue was obtained, without stromal cell or embryonic contamination, with the objective of identifying genes expressed exclusively in the luminal epithelium during the apposition stage of implantation. Total cellular RNA was then amplified to obtain aRNA for microarray hybridization to elucidate genes differentially expressed in the luminal epithelium of implantation (IM) sites compared to the interimplantation (INTER) sites (no blastocyst implantation).3


Method
SLIDE PREPARATION AND STAINING

1. Freshly frozen tissues were cut into 5 m thick sections and placed ont
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