Bioengineering anembryonic human trophoblast vesicles.
BACKGROUND
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.
Departments of Obstetrics and Gynecology, Warren Alpert Medical School at Brown University, Providence, RI 02067, USA. JRobins@wihri.org
Jared C
JC
Robins
Jeffrey R
JR
Morgan
Paula
P
Krueger
Sandra A
SA
Carson
D001485:Basement Membrane / Q000166:cytology; D057005:Bioengineering / Q000379:methods*; D019149:Bioreactors; D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D005353:Fibronectins; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D011247:Pregnancy; D012685:Sepharose; D018874:Spheroids, Cellular / Q000166:cytology* / Q000648:ultrastructure; D014327:Trophoblasts / Q000166:cytology* / Q000502:physiology*
D005353:Fibronectins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D012685:Sepharose