title: Establishment of trophoblast stem cell lines from somatic cell nuclear-transferred embryos.
abstract: Placental abnormalities occur frequently in cloned animals. Here, we attempted to isolate trophoblast stem (TS) cells from mouse blastocysts produced by somatic cell nuclear transfer (NT) at the blastocyst stage (NT blastocysts). Despite the predicted deficiency of the trophoblast cell lineage, we succeeded in isolating cell colonies with typical morphology of TS cells and cell lines from the NT blastocysts (ntTS cell lines) with efficiency as high as that from native blastocysts. The established 10 ntTS cell lines could be maintained in the undifferentiated state and induced to differentiate into several trophoblast subtypes in vitro. A comprehensive analysis of the transcriptional and epigenetic traits demonstrated that ntTS cells were indistinguishable from control TS cells. In addition, ntTS cells contributed exclusively to the placenta and survived until term in chimeras, indicating that ntTS cells have developmental potential as stem cells. Taken together, our data show that NT blastocysts contain cells that can produce TS cells in culture, suggesting that proper commitment to the trophoblast cell lineage in NT embryos occurs by the blastocyst stage.
authors.affiliation: Laboratory of Cellular Biochemistry, Animal Resource Sciences/Veterinary Medical Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Mayumi
authors.initials: M
authors.lastname: Oda
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tanaka
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yamazaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ohta
authors.forename: Misa
authors.initials: M
authors.lastname: Iwatani
authors.forename: Masako
authors.initials: M
authors.lastname: Suzuki
authors.forename: Jun
authors.initials: J
authors.lastname: Ohgane
authors.forename: Naka
authors.initials: N
authors.lastname: Hattori
authors.forename: Ryuzo
authors.initials: R
authors.lastname: Yanagimachi
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Wakayama
authors.forename: Kunio
authors.initials: K
authors.lastname: Shiota
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D015152:Blotting, Northern; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D065150:Cellular Reprogramming; D004622:Embryo, Mammalian / Q000166:cytology* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D051379:Mice; D053652:Nuclear Transfer Techniques; D020411:Oligonucleotide Array Sequence Analysis; D032681:Spectral Karyotyping; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism
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