Formation of Bovine Placental Trophoblast Spheroids

Abstract

Introduction: In this study, we aimed to form spheroids with the bovine placental trophoblast cell line F3. Spheroids are 3-dimensional culture models which can be used to conduct versatile in vitro and in vivo experiments. Materials and Methods: The spheroids were generated using the hanging drop technique, 25% methocel and matrigel. The F3 spheroids were characterized morphologically by light microscopy and transmission (TEM) and scanning electron microscopy (SEM) and immunohistochemistry (ezrin, vimentin, cytokeratin, placental lactogen). The fluorescent dyes calcein and ethidium homodimer were used to determine the viability of the spheroidal F3 cells by immunofluorescence microscopy. Results: The cell line F3 only formed spheroids by the hanging drop technique when matrigel was added. The trophoblast spheroids were delimited and fully covered by extracellular matrix (light microscopy/TEM/SEM). Cells contributing to spheroids could not be discriminated from each other (light microscopy). The outer spheroidal layer consisted of cells which possessed an apical pole with microvilli that were directed to the outside (light microscopy/TEM). All of the spheroidal F3 cells expressed ezrin, vimentin and cytokeratin, but not placental lactogen. The spheroid core contained degenerating cells whilst the F3 cells of the outer rim were viable (TEM/immunofluorescence microscopy). Discussion: We have established a 3-dimensional spheroid model for the bovine placental trophoblast cell line F3. The developed culture model might prove valuable for future in vitro studies on the differentiation of bovine trophoblast cells.

Evolution of placental invasion and cancer metastasis are causally linked

“…Human ESFs were immortalized by the Charles Lockwood lab at Yale, and obtained from Gil Mor group 72 . F3 cells were previously established by Pfarrer group at the University of Veterinary Medicine, Hannover, Germany 73,74 . J3, HTR8/SVNeo, BeWo, and BJ5ta cells were obtained from ATCC.…”

Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

“…25,26 Microarray Analysis RNA isolation, microarray analysis, and SYBR green quantitative real-time PCR was carried out, as described in the Methods in the online-only Data Supplement.…”

The Placenta in Toxicology. Part I

“…Bovine midgestational placenta (Haeger et al 2011), rat placental tissue (GD 20), and human placental tissue at the end of pregnancy (39th week) were also included in the study and obtained at the Department of Anatomy, University of Veterinary Medicine Hannover, Hannover, Germany or at the Department of Obstetrics and Gynecology at the Justus Liebig University Giessen, Giessen, Germany. The collection of human tissue was approved by the local ethics committee and informed consent was given in all cases (Haeger et al 2011). Porcine placental samples (GD 112) were a kind gift from Vibeke Dantzer (Department of Basic Animal and Veterinary Sciences, Copenhagen University, Copenhagen, Denmark).…”

“…In view of antibody application on nonhuman target tissues, we furthermore have also used marmoset organs as additional controls. Specificity of further antibodies used in this article have been described (Buse, Zoeller, and van Esch 2008; Haeger et al 2011; Martin and Buse 2008; van Esch et al 2008).…”