Placenta-on-a-chip model for assessing the transport and toxicity of xenobiotics in vitro
Objectives: The study of the transport and toxicity of xenobiotics in women is limited for ethical reasons. Ex vivo placenta models have high variability and low success rates. Animal models in vivo differ from a human in anatomy, genotype, and proteome. The placenta-on-a-chip model is a compromise. We studied the components of the FAC chemotherapy regimen for breast cancer in this model.
Methods: BeWo b30 cell line was grown in the DMEM with L-glutamine, 4.5 g glucose/l and Earle’s salts containing 10% FBS, 1x MEM NEAA, 100 U/ml penicillin and 100 μg/ml streptomycin in inserts cut from 96-well Transwell plate and placed in a microfluidic chip. Cells were seeded with a density of 10,000 cells per insert. After 7 days, 5-fluorouracil (25 μg/ml), doxorubicin (50 μg/ml), cyclophosphamide (150 μg/ml), or all three drugs were added to the cells for 1 hour. Control cells were cultured in the presence of 0.05% DMSO. The impedance spectrum was measured before and 1 and 24 hours after the addition of the drug. The concentration of the drug was determined by HPLC-MS/MS. Cell viability was assessed using the CellTiter-Blue Assay.
Results: After 1 h incubation with drugs, TEER decreased in experiment and control groups from an average of 90 to 25 Ω∙cm2, and after 24 h TEER was 67.3±17.9 Ω∙cm2 for control, 67.8±16.4 Ω∙cm2 for cyclophosphamide, 90.0±20.1 Ω∙cm2 for 5-fluorouracil, and decreased to the background for doxorubicin and drug mixture. Cell viability did not differ significantly between the control, 5-fluorouracil, and cyclophosphamide groups, but decreased to 40±9% of the control when exposed to doxorubicin and drug mixture. The placenta-on-a-chip model transported the drugs from the apical to the basolateral side.
Conclusion: The developed placenta-on-a-chip model is suitable for assessing the transport and toxicity of xenobiotics in vitro.
This work is supported by the Ministry of Science and Higher Education of the Russian Federation, project: 14.588.21.0007, unique id: RFMEFI58817X0007.