IMPLANTATION AND PREGNANCY - A
Monday, August 2, 2004
10:30 AM–12:30 PM
(262) COMPLIMENTARY DNA ARRAY ANALYSIS OF A CO-CULTURE MODEL OF IMPLANTATION: A COMPARISON OF SAM AND ARRAYINFORMATICS.
Walley, Rana1, Hines, Randall, 1 University of Mississippi Medical Center, Jackson, MS
ABSTRACT- Genetic changes in the human endometrium that result in implantation of the embryo are poorly understood. Using cDNA array analysis, we have developed an endometrial-trophoblast co-culture model to define important alterations in gene expression that may be associated with implantation. Human endometrium was obtained by biopsy from fertile, cycling women in the luteal phase (n=5). Endometrium was decidualized with progesterone and estradiol, and rising prolactin concentrations in the culture supernatant confirmed viability. Control endometrium was cultured under identical conditions to experimental endometrium with the exception of a 4 hour co-culture against BeWo cells in the experimental samples. The co-culture uses a 3m semi-permeable membrane which allows only soluble factors from the trophoblast to contact the endometrium. RNA was extracted from control and experimental endometrium and hybridized to single spotted 19K human arrays from the Microarry Centre in Toronto, Ontario, Canada. Microarray images were obtained using the PerkinElmer ScanArray Express. Digital data was analyzed from twenty separate cDNA arrays using both Significance Analysis of Microarrays (SAM) from Stanford University and ArrayInformatics 2.1 from Perkin Elmer. A total of 19200 genes were compared between control and experimental endometrium. SAM yielded only down regulation while ArrayInformatics showed both up and down regulation. Results will be confirmed with quantitative RT-PCR.
KEY WORDS: trophoblast, gene expression, microarray, co-culture