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Bioengineering Anembryonic Human Trophoblast Vesicles

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Abstract

Introduction

Trophoblast cells in vivo form a 3-dimensional structure that promotes complex cell-to-cell interactions that cannot be studied with traditional monolayer culture. We describe a 3-dimensional trophoblast bioreactor to study cellular interactions.

Methods

Nonadhesive agarose hydrogels were cast from molds using computer-assisted prototyping. Trophoblast cells were seeded into the gels for 10 days. Morphology, viability, and vesicle behavior were assessed.

Results

Trophoblast cells formed uniform spheroids. Serial sectioning on days 3, 7, and 10 revealed central vacuolization with a consistent outer rim 12.3-µ thick. The vesicle configuration has been confirmed with confocal imaging. Electron Microscopic (EM) imaging revealed its ultrastructure. The vesicles migrate across a fibronectin-coated surface and invaded basement membrane.

Conclusions

Trophoblast cells cultured in a novel substrate-free 3-dimensional system form trophoblast vesicles. This new cell culture technique allows us to better study placental cell-to-cell interactions with the potential of forming microtissues.

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Authors and Affiliations

  1. Departments of Obstetrics and Gynecology, Warren Alpert School of Medicine at Brown University, Women and Infants Hospital of Rhode Island, 101 Dudley Street, Providence, RI, 02067, USA

    Jared C. Robins MD, Paula Krueger BS & Sandra A. Carson MD

  2. Department of Molecular Pharmacology, Physiology and Biotechnology and Center for Biomedical Engineering, Warren Alpert Medical School at Brown University, Providence, RI, USA

    Jeffrey R. Morgan PhD

Authors
  1. Jared C. Robins MD
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  2. Jeffrey R. Morgan PhD
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  3. Paula Krueger BS
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  4. Sandra A. Carson MD
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Correspondence to Jared C. Robins MD.

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Robins, J.C., Morgan, J.R., Krueger, P. et al. Bioengineering Anembryonic Human Trophoblast Vesicles. Reprod. Sci. 18, 128–135 (2011). https://doi.org/10.1177/1933719110381923

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  • DOI: https://doi.org/10.1177/1933719110381923

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