Journal
INTERNATIONAL JOURNAL OF MEDICAL SCIENCES
Volume 18, Issue 14, Pages 3271-3279Publisher
IVYSPRING INT PUBL
DOI: 10.7150/ijms.61720
Keywords
AU-Siriraj; Cell polarization; Fetal bovine serum; Protein supplement; Renal tubular cell; Transepithelial electrical resistance
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Funding
- Office of National Higher Education Science Research and Innovation Policy Council (NXPO) through PMU-B
- Thailand Research Fund [IRN60W0004]
- Faculty of Medicine Siriraj Hospital
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The study found that adding 2.5% fetal bovine serum to artificial urine can prolong the deformation time of MDCK renal tubular cells and enhance polarization, but may increase cell death rate. Therefore, the use of AU-Siriraj+2.5% FBS is recommended for studying cell biology and function.
Several artificial urine (AU) formulas have been developed to mimic the normal urine. Most of them are protein-free, particularly when secreted proteins (secretome) is to be analyzed. However, the normal urine actually contains a tiny amount of proteins. We hypothesized that urinary proteins at physiologic level play a role in preservation of renal cell biology and function. This study evaluated the effects from supplementation of 0-10% fetal bovine serum (FBS) into the well-established AU-Siriraj protocol on MDCK renal tubular cells. Time to deformation (TD) was reduced by both native urine and AU-Siriraj without/with FBS compared with complete culture medium (control). Among the native urine and AU-Siriraj without/with FBS, the cells in AU-Siriraj+2.5% FBS had the longest TD. Supplementation of FBS increased cell death in a dose-dependent manner (but still <10%). Transepithelial electrical resistance (TER) of the polarized cells in the native urine was comparable to the control, whereas that of the cells in AU-Siriraj+2.5% FBS had the highest TER. These data indicate that supplementation of 2.5% FBS into AU-Siriraj can prolong time to deformation and enhance polarization of renal tubular cells. Therefore, AU-Siriraj+2.5% FBS is highly recommended for in vitro study of cell biology and function (when secretome is not subjected to analysis).
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