Publication: Optimization of artificial urine formula for in vitro cellular study compared with native urine
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Issued Date
2021-01-01
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14491907
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2-s2.0-85113777524
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Mahidol University
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SCOPUS
Bibliographic Citation
International Journal of Medical Sciences. Vol.18, No.14 (2021), 3271-3279
Suggested Citation
Kanyarat Sueksakit, Visith Thongboonkerd Optimization of artificial urine formula for in vitro cellular study compared with native urine. International Journal of Medical Sciences. Vol.18, No.14 (2021), 3271-3279. doi:10.7150/IJMS.61720 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/78634
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Title
Optimization of artificial urine formula for in vitro cellular study compared with native urine
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Abstract
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).