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Hum. Reprod. Advance Access published online on June 19, 2007
Human Reproduction, doi:10.1093/humrep/dem144
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© The Author 2007. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

The isolation and characterization of a population of extravillous trophoblast progenitors from first trimester human placenta

Joanna L. James1, Peter R. Stone and Lawrence W. Chamley

Department of Obstetrics and Gynecology, University of Auckland, Private Bag 92019, Auckland 1001, New Zealand

1 Correspondence address. Tel: +649 3737599; Fax: +649 3035969; E-mail: j.james{at}auckland.ac.nz

BACKGROUND: It is widely accepted that most if not all villous cytotrophoblasts from term placentae are committed to differentiate into syncytiotrophoblast, but that early in gestation villous cytotrophoblasts are bipotential and capable of differentiating into either extravillous trophoblasts (EVTs) or syncytiotrophoblast. In contrast, our previous work has suggested that two separate populations of cytotrophoblast exist in the first trimester, one committed to EVT differentiation and the other to syncytiotrophoblast differentiation. In this work, we have isolated and characterized the population of ‘EVT progenitors’.

METHODS: First trimester villous explants were cultured for 10 days then subjected to sequential trypsinization. Viable cells that adhered to Matrigel following trypsinization were cultured for up to 5 days and characterized by immunohistochemistry.

RESULTS: The isolation protocol yielded >90% cytokeratin positive trophoblasts, which expressed markers characteristic of EVT progenitors. Over 5 days of culture, these isolated putative EVT progenitors did not syncytialize, but ~20% differentiated into HLA-G positive EVTs.

CONCULSIONS: It is likely that the isolated putative EVT progenitors are the population of EVT progenitors previously identified in vivo. The characteristics of these isolated putative EVT progenitors provides further evidence for separate progenitors of EVT and syncytiotrophoblast in the first trimester.

Key words: trophoblast/placenta/differentiation/cytotrophoblast/extravillous trophoblast

Submitted on February 25, 2007; resubmitted on April 22, 2007; accepted on May 1, 2007.


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