Journal ArticleDOI
Purification, Characterization, and in vitro Differentiation of Cytotrophoblasts from Human Term Placentae*
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It is concluded that human cytotrophoblasts differentiate in culture and fuse to form functional syncytiotrophobic cells, similar to that of intact term placentae.Abstract:
Highly purified functional cytotrophoblasts have been prepared from human term placentae by adding a Percoll gradient centrifugation step to a standard trypsin-DNase dispersion method. The isolated mononuclear trophoblasts averaged 10 microns in diameter, with occasional cells measuring up to 20-30 microns. Viability was greater than 90%. Transmission electron microscopy revealed that the cells had fine structural features typical of trophoblasts. In contrast to syncytial trophoblasts of intact term placentae, these cells did not stain for hCG, human placental lactogen, pregnancy-specific beta 1-glycoprotein or low mol wt cytokeratins by immunoperoxidase methods. Endothelial cells, fibroblasts, or macrophages did not contaminate the purified cytotrophoblasts, as evidenced by the lack of immunoperoxidase staining with antibodies against vimentin or alpha 1-antichymotrypsin. The cells produced progesterone (1 ng/10(6) cells . 4 h), and progesterone synthesis was stimulated up to 8-fold in the presence of 25-hydroxycholesterol (20 micrograms/ml). They also produced estrogens (1360 pg/10(6) cells . 4 h) when supplied with androstenedione (1 ng/ml) as a precursor. When placed in culture, the cytotrophoblasts consistently formed aggregates, which subsequently transformed into syncytia within 24-48 h after plating. Time lapse cinematography revealed that this process occurred by cell fusion. The presumptive syncytial groups were proven to be true syncytia by microinjection of fluorescently labeled alpha-actinin, which diffused completely throughout the syncytial cytoplasm within 30 min. Immunoperoxidase staining of cultured trophoblasts between 3.5 and 72 h after plating revealed a progressive increase in cytoplasmic pregnancy-specific beta 1-glycoprotein, hCG, and human placental lactogen concomitant with increasing numbers of aggregates and syncytia. At all time points examined, occasional single cells positive for these markers were identified. RIA of the spent culture media for hCG revealed a significant increase in secreted hCG, paralleling the increase in hCG-positive cells and syncytia identified by immunoperoxidase methods. We conclude that human cytotrophoblasts differentiate in culture and fuse to form functional syncytiotrophoblasts.read more
Citations
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Journal ArticleDOI
A class I antigen, HLA-G, expressed in human trophoblasts.
Susan Kovats,Elliott K. Main,Clifford Librach,Marcia Stubblebine,Susan J. Fisher,Robert DeMars +5 more
TL;DR: HLA-G is subject to both cell type-specific and developmental regulation and is expressed in early gestation human cytotrophoblasts and is present in choriocarcinoma cell lines studied.
Journal ArticleDOI
92-kD type IV collagenase mediates invasion of human cytotrophoblasts.
Clifford Librach,Zena Werb,Marilyn L. Fitzgerald,King Chiu,Nanci M. Corwin,Ruth A. Esteves,Damian Grobelny,Richard E. Galardy,Caroline H. Damsky,Susan J. Fisher +9 more
TL;DR: It is concluded that the 92-kD type IV collagenase is critical for cytotrophoblast invasion, whereas inhibitors of the plasminogen activator system had only a partial inhibitory effect.
Journal ArticleDOI
Direct evidence to support the role of HLA-G in protecting the fetus from maternal uterine natural killer cytolysis
Nathalie Rouas-Freiss,Rachel Marchal-Bras Gonçalves,Catherine Menier,Jean Dausset,Edgardo D. Carosella +4 more
TL;DR: Evidence is provided under physiological conditions that supports the role of HLA-G in protecting cytotrophoblasts against natural killer cytolysis in 6 semiallogenic combinations of maternal uterine NK cells and their own trophoblast counterparts, and that their interactions contribute to the survival of the fetal semiallograft by confering immunological tolerance to its tissues.
Journal ArticleDOI
Obesity in pregnancy stimulates macrophage accumulation and inflammation in the placenta.
Challier Jc,Subhabrata Basu,T. Bintein,Judi Minium,K. Hotmire,Patrick M. Catalano,S. Hauguel-de Mouzon +6 more
TL;DR: The chronic inflammation state of pre-gravid obesity is extending to in utero life with accumulation of a heterogeneous macrophage population and pro-inflammatory mediators in the placenta.
Journal ArticleDOI
Transcytosis: Crossing Cellular Barriers
Pamela L. Tuma,Ann L. Hubbard +1 more
TL;DR: The authors' knowledge of the different cell types using transcytosis in vivo, the variety of cargo moved, and the diverse pathways for delivering that cargo are summarized.
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