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This Article Full Text Full Text (PDF) All Versions of this Article: 73/1/149    most recent biolreprod.104.037150v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wang, L. Articles by Tian, X. C. Search for Related Content PubMed PubMed Citation Articles by Wang, L. Articles by Tian, X. C. Agricola Articles by Wang, L. Articles by Tian, X. C.

Generation and Characterization of Pluripotent Stem Cells from Cloned Bovine Embryos¹

Institute of Zoology, Chinese Academy of Sciences,³ Beijing 100080, People's Republic of China Center for Regenerative Biology/Department of Animal Science,⁴ University of Connecticut, Storrs, Connecticut 06269

Bovine embryonic stem (ES) cell lines reported to date vary in morphology and marker expression (e.g., alkaline phosphatase [ALPL], stage-specific embryonic antigen 4 [SSEA4], and OCT4) that normally are associated with the undifferentiated, pluripotent state. These observations suggest that the proper experimental conditions for consistently producing bovine ES cells have not been identified. Here, we report three bovine ES cell lines, one from in vitro-fertilized and two from nuclear transfer embryos. These bovine ES cells grew in large, multicellular colonies resembling the mouse ES and embryonic germ (EG) cells and human EG cells. Throughout the culture period, most of the cells within the colonies stained positive for ALPL and the cell surface markers SSEA4 and OCT4. The staining patterns of nuclear transfer ES cells were identical to those of the blastocysts generated in vitro yet different from most previously reported bovine ES cell lines, which were either negative or not detected. After undifferentiated culture for more than 1 yr, these cells maintained the ability to differentiate into embryoid bodies and derivatives of all three EG layers, thus demonstrating their pluripotency. However, unlike the mouse and human ES cells, following treatment with trypsin, type IV collagenase, or protease E, our bovine ES cells failed to self-renew and became spontaneously differentiated. Presumably, this resulted from an interruption of the self-renewal pathway. In summary, we generated pluripotent bovine ES cells with morphology similar to those of established ES cells in humans and mice as well as marker-staining patterns identical to those of the bovine blastocysts.

embryo, in vitro fertilization

² Correspondence: X. Cindy Tian, Center for Regenerative Biology, University of Connecticut, 1392 Storrs Road, U-4243, Storrs, CT 06269. FAX: 860 486 8809; xiuchun.tian{at}uconn.edu

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