000149099 001__ 149099
000149099 005__ 20250124213232.0
000149099 0247_ $$2doi$$a10.1177/0391398819866458
000149099 0248_ $$2sideral$$a112976
000149099 037__ $$aART-2020-112976
000149099 041__ $$aeng
000149099 100__ $$aSanchez-Romero, Natalia
000149099 245__ $$aA simple method for the isolation and detailed characterization of primary human proximal tubule cells for renal replacement therapy
000149099 260__ $$c2020
000149099 5060_ $$aAccess copy available to the general public$$fUnrestricted
000149099 5203_ $$aThe main physiological functions of renal proximal tubule cells in vivo are reabsorption of essential nutrients from the glomerular filtrate and secretion of waste products and xenobiotics into urine. Currently, there are several established cell lines of human origin available as in vitro models of proximal tubule. However, these cells appeared to be limited in their biological relevance, because essential characteristics of the original tissue are lost once the cells are cultured. As a consequence of these limitations, primary human proximal tubule cells constitute a suitable and a biologically more relevant in vitro model to study this specific segment of the nephron and therefore, these cells can play an important role in renal regenerative medicine applications. Here, we describe a protocol to isolate proximal tubule cells from human nephrectomies. We explain the steps performed for an in-depth characterization of the cells, including the study of markers from others segments of the nephron, with the goal to determine the purity of the culture and the stability of proteins, enzymes, and transporters along time. The human proximal tubule cells isolated and used throughout this study showed many proximal tubule characteristics, including monolayer organization, cell polarization with the expression of tight junctions and primary cilia, expression of proximal tubule-specific proteins, such as megalin and sodium/glucose cotransporter 2, among others. The cells also expressed enzymatic activity for dipeptidyl peptidase IV, as well as for gamma glutamyl transferase 1, and expressed transporter activity for organic anion transporter 1, P-glycoprotein, multidrug resistance proteins, and breast cancer resistance protein. In conclusion, characterization of our cells confirmed presence of putative proximal tubule markers and the functional expression of multiple endogenous organic ion transporters mimicking renal reabsorption and excretion. These findings can constitute a valuable tool in the development of bioartificial kidney devices.
000149099 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/BES-2012-059562$$9info:eu-repo/grantAgreement/ES/MINECO/DPI2011-28262-C04-02
000149099 540__ $$9info:eu-repo/semantics/openAccess$$aAll rights reserved$$uhttp://www.europeana.eu/rights/rr-f/
000149099 590__ $$a1.595$$b2020
000149099 591__ $$aTRANSPLANTATION$$b20 / 25 = 0.8$$c2020$$dQ4$$eT3
000149099 591__ $$aENGINEERING, BIOMEDICAL$$b75 / 89 = 0.843$$c2020$$dQ4$$eT3
000149099 592__ $$a0.413$$b2020
000149099 593__ $$aBioengineering$$c2020$$dQ3
000149099 593__ $$aMedicine (miscellaneous)$$c2020$$dQ3
000149099 593__ $$aBiomedical Engineering$$c2020$$dQ3
000149099 593__ $$aBiomaterials$$c2020$$dQ3
000149099 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000149099 700__ $$aMartinez-Gimeno, Laura
000149099 700__ $$aGaetano-Pinto, Pedro
000149099 700__ $$0(orcid)0000-0003-2627-5953$$aSaez, Berta
000149099 700__ $$0(orcid)0000-0002-2328-9236$$aSanchez-Zalabardo, José Manuel
000149099 700__ $$aMasereeuw, Rosalinde
000149099 700__ $$0(orcid)0000-0002-6043-4869$$aGimenez, Ignacio$$uUniversidad de Zaragoza
000149099 7102_ $$11012$$2410$$aUniversidad de Zaragoza$$bDpto. Farmac.Fisiol.y Med.L.F.$$cÁrea Fisiología
000149099 773__ $$g43, 1 (2020), 45–57$$pInt. j. artif. organs$$tINTERNATIONAL JOURNAL OF ARTIFICIAL ORGANS$$x0391-3988
000149099 8564_ $$s2552124$$uhttps://zaguan.unizar.es/record/149099/files/texto_completo.pdf$$yPostprint
000149099 8564_ $$s2169770$$uhttps://zaguan.unizar.es/record/149099/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000149099 909CO $$ooai:zaguan.unizar.es:149099$$particulos$$pdriver
000149099 951__ $$a2025-01-24-21:10:10
000149099 980__ $$aARTICLE