Three-dimensional growth of extravillous cytotrophoblasts promotes differentiation and invasion

• Published in: Placenta (Eastbourne) 2005-11, Vol.26 (10), p.709-720
• Main Authors: LaMarca, H.L., Ott, C.M., Höner zu Bentrup, K., LeBlanc, C.L., Pierson, D.L., Nelson, A.B., Scandurro, A.B., Whitley, G. St. J., Nickerson, C.A., Morris, C.A.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Bioreactors; Blotting, Western; Cell Aggregation - physiology; Cell Differentiation - physiology; Cell Growth Processes - physiology; Cell Line; Cytotrophoblast; Differentiation; Embryology: invertebrates and vertebrates. Teratology; Female; Fundamental and applied biological sciences. Psychology; Humans; Invasion; L-Selectin - biosynthesis; L-Selectin - genetics; Matrix metalloproteinase; Matrix Metalloproteinases - genetics; Matrix Metalloproteinases - metabolism; Matrix Metalloproteinases - secretion; Microscopy, Electron, Scanning; Microscopy, Fluorescence; Placentation; Placentation - physiology; Platelet Endothelial Cell Adhesion Molecule-1 - biosynthesis; Platelet Endothelial Cell Adhesion Molecule-1 - genetics; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Rotating wall vessel; Three-dimensional; Trophoblasts - cytology; Trophoblasts - enzymology; Trophoblasts - secretion; Trophoblasts - ultrastructure; Urokinase-Type Plasminogen Activator - genetics; Urokinase-Type Plasminogen Activator - metabolism; Urokinase-Type Plasminogen Activator - secretion
• ISSN: 0143-4004; 1532-3102
• DOI: 10.1016/j.placenta.2004.11.003

Human trophoblast research relies on a combination of in vitro models, including isolated primary cultures, explant cultures, and trophoblast cell lines. In the present study, we have utilized the rotating wall vessel (RWV) bioreactor to generate a three-dimensional (3-D) model of human placentation for the study of cytotrophoblast (CTB) invasion. The RWV supported the growth of the human CTB cell line SGHPL-4 and allowed for the formation of complex, multilayered 3-D aggregates that were morphologically, phenotypically, and functionally distinct from SGHPL-4 monolayers. The cells cultured three-dimensionally differentiated into an aggressively invasive cell population characterized by the upregulation of matrix metalloproteinase-2 (MMP-2), MMP-3, MMP-9 and urokinase-type plasminogen activator (uPA) secretion and activation. Microarray analysis of the 3-D and 2-D cultured cells revealed increased expression in the 3-D cells of various genes that are known mediators of invasion, including MT1-MMP, PECAM-1 and L-selectin, as well as genes not previously associated with CTB differentiation such as MMP-13 and MT5-MMP. These results were verified by quantitative real-time PCR. These findings suggest that when cultured in 3-D, SGHPL-4 cells closely mimic differentiating in utero CTBs, providing a novel approach for the in vitro study of the molecular mechanisms that regulate CTB differentiation and invasion.