International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)


I11. Genome-wide Expression Profiling of Embryonic Stem Cells and Trophoblast Stem Cells with NIA Mouse 15k cDNA Microarray

Tetsuya S. Tanaka1, Saied A. Jaradat1, Tilo Kunath2, Yulan Piao1, Janet Rossant2 and Minoru S.H. Ko1
1Laboratory of Genetics, National Institute on Aging, NIH, Baltimore, MD 21224, USA.
2Samuel Lunenfeld Research Institute, Mount Sinai Hospital, University of Toronto,Toronto, Ontario M5G 1X5, Canada

The first cell differentiation in placental mammals gives rise to two different cell types; Inner cell mass (ICM) that develops into the embryo proper, and Trophectoderm (TE) that differentiates into placental trophoblast, cells indispensable to sustain embryonic development in utero. To characterize this differentiation process, we have employed the method of global gene expression profiling on cDNA microarrays. As a first step, we took advantage of cell culture systems that provide enough RNA to probe microarrays. Cultured in vitro, the ICM-derived embryonic stem (ES) cells and TE-derived trophoblast stem (TS) cells (Science 282: 2072-5, 1998) provided probes that were hybridized to NIA mouse 15K cDNA microarrays (Proc. Natl. Acad. Sci. USA, in press.; http://lgsun.grc.nia.nih.gov/), containing ~15,000 unique genes collected from early embryos, including preimplantation embryos. Experiments were done in triplicate to obtain statistically significant results. Approximately 1000 genes showed statistically significantly different expression levels between ES and TS cells at the 5% confidence level. As expected, many TS cell-specific genes turn out to be expressed by independent assays of mid-gestation placenta. For example, EndoA, TGase and Pem are expressed highly in both TS cells and mid-gestation placenta at the 5% confidence level. On the other hand, PSX, PL-I, eHAND and PRL, which are expressed highly in placenta, do not show statistically significant expression in TS cells. This may imply that EndoA, TGase and Pem are expressed in early stage of the extraembryonic lineage development. From this gene collection, one of the unknown genes with classic Zinc finger domain and mapped to the proximal region of Chromosome 2 (D2Wsu23e), is currently being analyzed more in detail by in situ hybridization and RT-PCR.


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