Energy metabolism and glycolysis in human placental trophoblast cells during differentiation.

Energy metabolism and glycolysis of normal human term placental trophoblast in two-sided culture was investigated during differentiation from cytotrophoblast to syncytiotrophoblast, because glycogen metabolism is abnormal in several trophoblast related pregnancy diseases, including pre-eclampsia. After initial recovery of energy and cytoplasmic NADH/NAD+ redox by 24 h of culture, measures of cellular energy state, [ATP], [ADP], [ATP]/[ADP] ratio, ([ATP] + [ADP] + [AMP]), [ATP]/([ATP] + [ADP] + [AMP]) and energy charge remained essentially constant until 72 h, despite periods of increased energy turnover. At 24 h there was a burst of glycogenolysis, and glycolysis indicated by increased lactate production, which coincided with formation of syncytium.

Human fetal endothelial cells acquire zinc(II) from both the protein bound and nonprotein bound pools in serum.

To help determine physiologically important routes by which zinc (Zn) is acquired by human fetal vascular endothelium, the authors incubated cultured umbilical vein endothelial cells with 65Zn(II)-tracer labeled human fetal whole serum, ultrafiltrate (containing low molecular mass serum zinc complexes), and dialyzed serum (containing protein-bound zinc). Zinc from whole serum and from both serum fractions entered a rapidly labeled cellular compartment, removable by edetic acid (EDTA), representing Zn bound to the outside cell surface, and accumulatively, an EDTA-resistant compartment-probably largely internalized Zn. Entry of Zn into the EDTA-resistant pool from both serum fractions was strongly temperature-dependent, and was not via the EDTA-sensitive pool.

Culture of syncytiotrophoblast for the study of human placental transfer. Part II: Production, culture and use of syncytiotrophoblast.

The conditions necessary for producing syncytical syncytiotrophoblast are examined. Tissue disaggregation conditions, culture media composition, different extracellular matrices and the influence of placental gestational age are all assessed. The importance of evaluating the biochemical and functional differentiational state of the cells is also stressed.

Culture of syncytiotrophoblast for the study of human placental transfer. Part I: Isolation and purification of cytotrophoblast.

Criteria for a successful model for the study of trans-syncytiotrophoblast transfer include isolating substantially pure trophoblast cells from placental villous tissue, and obtaining from them phenotypical villous syncytial syncytiotrophoblast during culture. For studies involving the basal membrane, including overall transfer, basal uptake and output, and controls acting at the basal membrane, a two-sided model is required with a separate compartment of culture medium in contact with the basal cell surface. All current methods of isolating cytotrophoblast, the precursor of syncytiotrophoblast, derive from the original tissue trypsinization method of Thiede (1960), which produces cultures of villous cytotrophoblast cells contaminated with other placental cell types.