Hyperglycaemia in vitro alters the proliferation and mitochondrial activity of the choriocarcinoma cell lines BeWo, JAR and JEG-3 as models for human first-trimester trophoblast.
OBJECTIVE
Early intrauterine growth delay in diabetes could be caused by a reduced growth of the placenta. Our study investigates whether hyperglycaemia in vitro reduces trophoblast proliferation.


METHODS
First-trimester trophoblast cell models (BeWo, JAR and JEG-3 choriocarcinoma cells) were cultured for 24 and 48 h with 5.5 mmol/l D-glucose, 25 mmol/1 D-glucose (hyperglycaemia) and with an osmotic control. Cell number, total protein and nucleic acid content and mitochondrial activity (tetrazolium salt assay) were measured, the cell cycle analysed (FACS, cyclin B1 levels) and apoptosis (Annexin-V) measured.


RESULTS
In BeWo cells hyperglycaemia reduced cell number, protein, nucleic acid and cyclin B1 levels. The reduced G2/M and increased G0/G1 population after 24 h reflects growth arrest at G0/G1. In JAR cells after 24 h the population was arrested in G0/G1, whereas after 48 h the G0/G1 block was abrogated and the cells were arrested at G2/M. The net effect was an unchanged cell number. In JEG-3 cells hyperglycaemia resulted in fewer cells after 24 h but not after 48 h indicating some adaptation. Mitochondrial activity was either stimulated (BeWo) or reduced (JAR, JEG-3) under hyperglycaemia. Some of these effects were also induced by hyperosmolarity alone.


CONCLUSIONS
Hyperglycaemia has the potential to inhibit the proliferation of first-trimester trophoblast cell models. The mechanisms leading to growth arrest and to changes in mitochondrial activity are complex and depend on differentiation. We hypothesise a hyperglycaemia-induced impairment of placental growth in the first trimester of a poorly controlled diabetic pregnancy.
Clinic of Obstetrics and Gynaecology, Graz, Austria.
U
U
Weiss
M
M
Cervar
P
P
Puerstner
O
O
Schmut
J
J
Haas
R
R
Mauschitz
G
G
Arikan
G
G
Desoye
D017209:Apoptosis; D002452:Cell Count; D002453:Cell Cycle; D002455:Cell Division; D002470:Cell Survival; D002822:Choriocarcinoma / Q000473:pathology*; D019926:Cyclin B / Q000032:analysis; D056744:Cyclin B1; D005260:Female; D005434:Flow Cytometry; D005865:Gestational Age; D005947:Glucose / Q000008:administration & dosage; D006801:Humans; D006943:Hyperglycemia / Q000473:pathology*; D015151:Immunoblotting; D008928:Mitochondria / Q000473:pathology*; D008954:Models, Biological; D009994:Osmolar Concentration; D011247:Pregnancy; D011254:Pregnancy in Diabetics / Q000473:pathology; D014327:Trophoblasts / Q000502:physiology* / Q000648:ultrastructure; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000473:pathology*
C534699:CCNB1 protein, human; D019926:Cyclin B; D056744:Cyclin B1; D005947:Glucose