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Human Molecular Genetics Advance Access published online on February 12, 2004
Human Molecular Genetics, doi:10.1093/hmg/ddh088
© 2004 by Oxford University Press
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©2004 Oxford University Press

Article

Spontaneous differentiation of germ cells from human embryonic stem cells in vitro

Amander T. Clark 1, Megan S. Bodnar 2, Mark Fox 1, Ryan T. Rodriquez 2, Michael J. Abeyta 1, Meri T. Firpo 2*, and Renee A. Reijo Pera 1*

1 Center for Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, University of California at San Francisco, San Francisco, CA 94143-0556, USA; Departments of Physiology and Urology, and Programs in Human Genetics, Cancer Genetics, University of California at San Francisco, San Francisco, CA 94143-0556, USA; Development and Stem Cell Biology, University of California at San Francisco, San Francisco, CA 94143-0556, USA
2 Center for Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, University of California at San Francisco, San Francisco, CA 94143-0556, USA; Development and Stem Cell Biology, University of California at San Francisco, San Francisco, CA 94143-0556, USA

* To whom correspondence should be addressed. E-mail: reijo{at}itsa.ucsf.edu.


  Abstract

Little is known of molecular requirements for specification of human germ cells. However, it is likely that they are specified through the action of sequentially-expressed genes just as in model organisms. We sought to determine whether human embryonic stem (ES) cell lines, like those of mice, might be capable of forming germ cells in vitro. We compared transcriptional profiles of three pluripotent human ES cells to those of isolated inner cell mass (ICM) cells. We found that ICM cells expressed NANOS1, STELLAR, and OCT4, whereas, undifferentiated human ES cells expressed these genes along with the germ cell specific gene, DAZL. Upon ES cell differentiation into embryoid bodies (EBs), we observed a shift in expression from RNA and protein markers of immature germ cells to those indicative of mature germ cells including expression of VASA, BOL, SCP1, SCP3, GDF9 and TEKT1. Although ability to test the function of these putative VASA positive germ cells is limited, these results demonstrate that differentiation of human ES cells into EBs in vitro results in formation of cells that express markers specific to gonocytes.


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