Polarized Epithelial Cell Culture

Epithelial cells were plated in 0.4 μm 24-well plate Matrigel-coated transwell inserts (Corning Life Sciences, Tewksbury, MA) and grown in defined media consisting of DMEM/F12 (Thermo Fisher) supplemented with NuSerum (Corning Life Sciences), Hyclone Defined FBS (GE Life Sciences, Logan, UT), Penicillin–Streptomycin (Thermo Fisher), l-Glutamine (Thermo Fisher) and HEPES (GE Life Sciences) as previously described by us^{30 (link),31}. Growth on transwell inserts allows for the formation of distinct apical and basolateral compartments^{30 (link),31}. Cells were grown to confluence and allowed to polarize as determined by transepithelial resistance (TER) of greater than 1000 ohms per insert^{30 (link),31}. TER was measured using an EVOM electrode and Voltohmmeter (World Precision Instruments, Sarasota, FL). Only polarized preparations of epithelial cells above 1000 ohms were used in this study. 24 h before progestin treatment, cells were transferred to stripped media.

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Patel M.V., Rodriguez-Garcia M., Shen Z, & Wira C.R. (2021). Medroxyprogesterone acetate inhibits wound closure of human endometrial epithelial cells and stromal fibroblasts in vitro. Scientific Reports, 11, 23246.

Publication 2021

Matrigel-coated transwell inserts DMEM/F12 NuSerum Hyclone Defined FBS Penicillin–Streptomycin l-Glutamine HEPES EVOM electrode and Voltohmmeter

Cells Epithelial cells Hepes L glutamine Matrigel Penicillin Progestin Streptomycin

Corresponding Organization : Dartmouth College

Other organizations : Tufts University

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Variable analysis

independent variables

Progestin treatment

dependent variables

Transepithelial resistance (TER)

control variables

Epithelial cell type
Culture medium composition (DMEM/F12, NuSerum, Hyclone Defined FBS, Penicillin–Streptomycin, L-Glutamine, HEPES)
Culture format (Matrigel-coated transwell inserts)
Incubation duration (cells grown to confluence and allowed to polarize)
Polarization criteria (TER > 1000 ohms per insert)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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