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Optimization of artificial urine formula for in vitro cellular study compared with native urine

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 phys...

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Autores principales: Sueksakit, Kanyarat, Thongboonkerd, Visith
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Ivyspring International Publisher 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8364465/
https://www.ncbi.nlm.nih.gov/pubmed/34400896
http://dx.doi.org/10.7150/ijms.61720
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author Sueksakit, Kanyarat
Thongboonkerd, Visith
author_facet Sueksakit, Kanyarat
Thongboonkerd, Visith
author_sort Sueksakit, Kanyarat
collection PubMed
description 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 (T(D)) 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 T(D). 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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spelling pubmed-83644652021-08-15 Optimization of artificial urine formula for in vitro cellular study compared with native urine Sueksakit, Kanyarat Thongboonkerd, Visith Int J Med Sci Research Paper 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 (T(D)) 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 T(D). 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). Ivyspring International Publisher 2021-07-23 /pmc/articles/PMC8364465/ /pubmed/34400896 http://dx.doi.org/10.7150/ijms.61720 Text en © The author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions.
spellingShingle Research Paper
Sueksakit, Kanyarat
Thongboonkerd, Visith
Optimization of artificial urine formula for in vitro cellular study compared with native urine
title Optimization of artificial urine formula for in vitro cellular study compared with native urine
title_full Optimization of artificial urine formula for in vitro cellular study compared with native urine
title_fullStr Optimization of artificial urine formula for in vitro cellular study compared with native urine
title_full_unstemmed Optimization of artificial urine formula for in vitro cellular study compared with native urine
title_short Optimization of artificial urine formula for in vitro cellular study compared with native urine
title_sort optimization of artificial urine formula for in vitro cellular study compared with native urine
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8364465/
https://www.ncbi.nlm.nih.gov/pubmed/34400896
http://dx.doi.org/10.7150/ijms.61720
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