Table 1.

Influence of low O2 and HBEGF signaling on cytotrophoblast survival, proliferation and HBEGF production

Proliferation (Ki67 index)Cell death (TUNEL index)Cellular HBEGF (normalized) 2% O2Secreted HBEGF (normalized) 2% O2
TreatmentConcentrationAmbient O22% O2Ambient O22% O2
Vehicle11.4±1.041.8±2.54.75±0.554.50±0.23100±25100±26
CRM19710 μg/ml10.2±0.6337.6±2.83.94±0.439.86±0.41*0.943±0.34*ND
Anti-HBEGF10 μg/ml9.52±0.8234.1±2.793.93±0.6911.5±0.56*1.37±0.70*ND
Heparitinase0.1 U/ml3.66±0.3811.5±0.63*
Anti-HER1 and HER410 μg/ml each10.1±0.5237.3±2.344.85±0.3310.1±0.46*5.51±2.5*4.66±4.54*
Anti-HER110 μg/ml4.14±0.4957.0±3327.7±18.7*
Anti-HER410 μg/ml4.34±0.8151.6±2025.2±17.3*
Non-Immune IgG20 μg/ml5.10±0.575.24±0.40
  • Cytotrophoblast cells were cultured at the indicated O2 levels for either 4 (HBEGF measurement) or 8 (cell death and proliferation) hours. All treatments were initiated 30 minutes before the start of culture and continued for the remainder of the culture period. Cells and medium were collected and assayed for Ki67 expression, TUNEL and HBEGF, as described in the Materials and methods section. Mean±s.e.m. is shown for all measurements. HBEGF concentrations, determined by ELISA, were normalized as a percentage of the vehicle treatment.

    ND, not done because the presence of CRM197 or anti-HBEGF in medium interferes with the HBEGF ELISA.

  • * P<0.05, compared with vehicle at the same O2 level.