Culturing and Characterizing M-1 Kidney Cells

The M-1 cell line derived from the CD of mice^{28 (link)}, was purchased from American Type Culture Collection (ATCC, Manassas, VA). M-1 cells are composed by principal and intercalated cells, this cell model has been validated previously^{29 (link)–31 (link)}. Prior to the experiments, M-1 cells were maintained in Dulbecco′s modified Eagle’s medium (DMEM)/F-12 medium (LIFE TECHNOLOGIES, Carlsbad, CA) containing 10% fetal bovine serum (FBS), growth factors (transferrin, insulin, and sodium selenite at physiological concentrations), 100 nmol/L dexamethasone and 5 mM glucose. When the cells reached approximately 70–75% confluence, they were dissociated using trypsin and split into 4-wells chambers and dishes for the trafficking assays.

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Gogulamudi V.R., Arita D.Y., Bourgeois C.R., Jorgensen J., He J., Wimley W.C., Satou R., Gonzalez A.A, & Prieto M.C. (2021). High glucose induces trafficking of prorenin receptor and stimulates profibrotic factors in the collecting duct. Scientific Reports, 11, 13815.

Publication 2021

M-1 cell line DMEM)/F-12 medium

Assays Cells Dexamethasone Dishes Eagle Fetal bovine serum Glucose Growth factors Insulin M cells Physiological Sodium selenite Transferrin Trypsin

Corresponding Organization : Tulane University

Other organizations : Pontificia Universidad Católica de Valparaíso

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Variable analysis

independent variables

Cell culture conditions (DMEM/F-12 medium containing 10% FBS, growth factors, 100 nmol/L dexamethasone, and 5 mM glucose)
Cell confluence (70-75%)

dependent variables

Not explicitly mentioned

control variables

Cell line (M-1 cells derived from the CD of mice)
Cell composition (principal and intercalated cells)
Cell dissociation method (trypsin)

controls

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

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