A human cytotrophoblast-villous endothelium-fetal organ multi-cell model and the impact on gene and protein expression in placenta cytotrophoblast, fetal hepatocytes and fetal kidney epithelial cells

人滋养层细胞-绒毛内皮细胞-胎儿器官多细胞模型及其对胎盘滋养层细胞、胎肝细胞和胎肾上皮细胞基因和蛋白质表达的影响

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Abstract

INTRODUCTION: Appropriate fetal growth requires multi-directional, coordinated communication between maternal, placental, and fetal systems. Disruptions in these signaling arms can have deleterious consequences for fetal growth and lead to fetal developmental adaptations associated with short- and long-term morbidities. This proof-of-concept human cell model study aimed to identify the effects of altered trophoblast culture conditions and human insulin-like 1 growth factor (hIGF1) nanoparticle gene therapy on fetal liver hepatocytes and kidney epithelial cells. METHODS: We utilized human cell lines: BeWo choriocarcinoma cells (trophoblast), Human Placental Micro-Vascular Endothelial Cells, and WRL68 (hepatocytes) or HEK293T/17 (kidney epithelium), in a co-culture model designed to mimic cytotrophoblast-villous endothelium-fetal organ communication. RESULTS: Trophoblast stress response mechanisms were increased by culturing BeWo cells in growth media without fetal bovine serum (FBS). BeWo cells were also cultured without FBS and treated with a hIGF1 nanoparticle gene therapy which is known to mitigate cellular stress mechanisms. BeWo cells without FBS support had increased expression of cellular stress mechanisms but not when IGF1 was over-expressed with a transient hIGF1 nanoparticle gene therapy. BeWo cells without FBS and without FBS + hIGF1 nanoparticle gene therapy had increased expression of gluconeogenesis and glycolysis rate-limiting enzymes. Gene and protein expression in fetal liver and kidney cells was not impacted by increased trophoblast stress or hIGF1 nanoparticle gene therapy. DISCUSSION: Our data demonstrated that cytotrophoblast, cultured without FBS support, turn on mechanisms involved in glucose production. Whether this is reflected in vivo remains uninvestigated but may represent a placental compensation mechanism in complicated pregnancies.

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