Abstract
AimsThe purpose of this study was to determine the cell viability and cytotoxicity of various endocytosis and efflux inhibitors which can be used to determine transport and uptake mechanisms in the BeWo (b30 clone) human placental trophoblast cell line. Ethanol and dimethylsulfoxide (DMSO) were also studied since they are often used as cosolvents for administration of these inhibitors.MethodologyThe water-soluble tetrazolium-1 (WST-1) assay was used to quantify cell viability and the lactate dehydrogenase (LDH) assay was used to determine cytotoxicity.ResultsBy the WST-1 assay, reduced cell viability was observed following 4 hours of exposure to chlorpromazine (10 μg/mL), colchicine (1 mM), filipin (3 μg/mL), gentamicin (2 mM), GF120918 (1 μM), methyl-β-cyclodextrin (5 mM), and verapamil (100 μM). By the LDH assay, however, no cytotoxicity was observed after 4 hours of exposure to the aforementioned compounds. Amiloride (500 μM), ethanol (up to 0.1% v/v), and DMSO (up to 0.1% v/v) did not reduce cell viability nor induce cytotoxicity.ConclusionThis information is valuable when selecting potential inhibitors of endocytosis and efflux and the selection of time points for mechanistic studies.
Highlights
Pregnant women are often excluded from clinical trials, many medications are still prescribed during pregnancy
A number of inhibitors were investigated as potential candidates to elucidate mechanisms of endocytosis or efflux in placental trophoblast cells for drugs or drug carriers that could be used during pregnancy
Cells treated with 0.1% (v/v) Triton X-100 exhibited the lowest cell viability according to the water-soluble tetrazolium-1 (WST-1) assay (Fig. 1)
Summary
Pregnant women are often excluded from clinical trials, many medications are still prescribed during pregnancy. The placental transport of these drugs and their effects upon fetal health and development are of concern. The BeWo b30 cell line is a placental trophoblast cell line of human origin that can form confluent monolayers to study the transport and metabolism of drugs across the human placental barrier [2,3]. The effect of incubation time on the cytotoxicity of these inhibitors in BeWo cells has not been investigated previously to the best of our knowledge. If these inhibitors are toxic to cells, it could result in misinterpretation of the mechanistic data obtained. We examined the effect of those inhibitors on BeWo cells, as well as the effects of ethanol and DMSO, which are common cosolvents used to solubilize those and other compounds
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