Human cytotrophoblasts were isolated from three or four individual placentas per experiment, as previously described [55 (link)]. Primary cytotrophoblasts were cultured in DMEM GlutaMAX (Life Technologies) containing 10% fetal calf serum and 1% antibiotic-antimycotic on fibronectin (10 mg/mL; BD Biosciences, Franklin Lakes, New Jersey, United States) coated wells. Cells were plated and allowed to attach over 12–18 h before washing with dPBS (Life Technologies) to remove cell debris. Cells were cultured under 8% O2, 5% CO2 at 37 °C.
Antibiotic antimycotic on fibronectin
Manufactured by BD
Sourced in United States
Antibiotic-antimycotic on fibronectin is a laboratory reagent used to maintain sterile cell culture conditions. It contains a combination of antibiotics and antifungal agents that inhibit the growth of bacteria, yeasts, and fungi in cell culture media. The reagent is typically applied to the surface of cell culture vessels coated with the extracellular matrix protein fibronectin, which can promote cell attachment and growth.
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2 protocols using antibiotic antimycotic on fibronectin
1
Isolation and Culture of Human Cytotrophoblasts
2
Isolation and Culture of Human Cytotrophoblasts
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Human cytotrophoblasts were isolated from term, caesarean section placentas as previously described40 (link),41 (link). Primary cytotrophoblasts were cultured in DMEM high Glutamax (Life Technologies) containing 10% FCS and 1% antibiotic–antimycotic on fibronectin (10 ug/mL; BD Biosciences, New South Wales, Victoria) coated wells. Cells were plated and allowed to attach over 12–18 h before washing with dPBS (Life Technologies) to remove cellular debris. Primary cytotrophoblasts were maintained in a humidified incubator at 8% O2 (to simulate placental normoxia) or 1% O2 (to simulate placental hypoxia) and 5% CO2 for 24 h37 .
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