Expression of GFP under the control of the RNA helicase VASA permits fluorescence-activated cell sorting isolation of human primordial germ cells.

Stem Cells
Katarzyna TilgnerLyle Armstrong

Abstract

The isolation of significant numbers of human primordial germ cells at several developmental stages is important for investigations of the mechanisms by which they are able to undergo epigenetic reprogramming. Only small numbers of these cells can be obtained from embryos of appropriate developmental stages, so the differentiation of human embryonic stem cells is essential to obtain sufficient numbers of primordial germ cells to permit epigenetic examination. Despite progress in the enrichment of human primordial germ cells using fluorescence-activated cell sorting (FACS), there is still no definitive marker of the germ cell phenotype. Expression of the widely conserved RNA helicase VASA is restricted to germline cells, but in contrast to species such as Mus musculus in which reporter constructs expressing green fluorescent protein (GFP) under the control of a Vasa promoter have been developed, such reporter systems are lacking in human in vitro models. We report here the generation and characterization of human embryonic stem cell lines stably carrying a VASA-pEGFP-1 reporter construct that expresses GFP in a population of differentiating human embryonic stem cells that show expression of characteristic markers of primordial g...Continue Reading

References

Aug 11, 1994·Nucleic Acids Research·S J ClarkM Frommer
Jul 25, 2000·Mechanisms of Development·A K BraatD Zivkovic
Aug 2, 2000·Proceedings of the National Academy of Sciences of the United States of America·D H CastrillonC P Crum
Jul 28, 2001·Human Reproduction·L JohnsonR Yanagimachi
Sep 22, 2001·Cell Structure and Function·T NoceN Tsunekawa
Sep 19, 2002·Molecular Reproduction and Development·Abraham L Kierszenbaum
Feb 20, 2003·DNA and Cell Biology·Richárd Bártfai, László Orbán
Mar 27, 2003·Molecular Reproduction and Development·Shun SatoYasuhisa Matsui
May 6, 2003·Science·Karin HübnerHans R Schöler
Sep 25, 2003·Proceedings of the National Academy of Sciences of the United States of America·Yayoi ToyookaToshiaki Noce
Dec 4, 2003·Molecular Reproduction and Development·Terumasa KobayashiToshio Takeuchi
Feb 14, 2004·Human Molecular Genetics·Amander T ClarkRenee A Reijo Pera
May 3, 2005·Nature Genetics·Peter J Rugg-GunnRoger A Pedersen
Apr 6, 2006·European Journal of Cell Biology·Robert HockUlrich Scheer
Nov 17, 2006·Biology of Reproduction·Leanne S Sleer, Christopher C Taylor
May 8, 2007·Proceedings of the National Academy of Sciences of the United States of America·Frédéric ChalmelMichael Primig
May 10, 2007·Nature Protocols·Jason A WestNiels Geijsen
Jun 16, 2007·Frontiers in Bioscience : a Journal and Virtual Library·John Arne Dahl, Philippe Collas
Jun 28, 2007·Seminars in Reproductive Medicine·Ping Zheng, Jurrien Dean
Jan 17, 2008·Nature Reviews. Genetics·Hiroyuki Sasaki, Yasuhisa Matsui
Mar 8, 2008·Molecular Reproduction and Development·Takeo MinematsuMitsuru Naito
Sep 20, 2008·Stem Cells·Katarzyna TilgnerLyle Armstrong
Nov 19, 2008·Developmental Biology·Shuichi ToyodaJun-ichi Miyazaki

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Citations

Jul 24, 2012·Epigenetics & Chromatin·Richard B Meagher, Kristofer J Müssar
Dec 5, 2015·Stem Cells International·Aleksandar NikolicMiodrag Stojkovic
Apr 21, 2021·Reproductive Sciences·Sivanandane SittadjodyJames J Yoo
Jun 18, 2011·Stem Cells·C EguizabalJ C Izpisua Belmonte
Feb 11, 2012·Reproduction : the Official Journal of the Society for the Study of Fertility·Zachary Yu-Ching LinHideyuki Okano
Jun 5, 2012·Fertility and Sterility·Yohei HayashiShinya Yamanaka

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