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A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens

The kidney is a target organ for the toxicity of several xenobiotics and is also highly susceptible to the development of malignant tumors. In both cases, in vitro studies provide insight to cellular damage, and represent adequate models to study either the mechanisms underlying the toxic effects of...

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Autores principales: Valente, Maria João, Henrique, Rui, Costa, Vera L., Jerónimo, Carmen, Carvalho, Félix, Bastos, Maria L., de Pinho, Paula Guedes, Carvalho, Márcia
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3087760/
https://www.ncbi.nlm.nih.gov/pubmed/21573239
http://dx.doi.org/10.1371/journal.pone.0019337
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author Valente, Maria João
Henrique, Rui
Costa, Vera L.
Jerónimo, Carmen
Carvalho, Félix
Bastos, Maria L.
de Pinho, Paula Guedes
Carvalho, Márcia
author_facet Valente, Maria João
Henrique, Rui
Costa, Vera L.
Jerónimo, Carmen
Carvalho, Félix
Bastos, Maria L.
de Pinho, Paula Guedes
Carvalho, Márcia
author_sort Valente, Maria João
collection PubMed
description The kidney is a target organ for the toxicity of several xenobiotics and is also highly susceptible to the development of malignant tumors. In both cases, in vitro studies provide insight to cellular damage, and represent adequate models to study either the mechanisms underlying the toxic effects of several nephrotoxicants or therapeutic approaches in renal cancer. The development of efficient methods for the establishment of human normal and tumor renal cell models is hence crucial. In this study, a technically simple and rapid protocol for the isolation and culture of human proximal tubular epithelial cells and human renal tumor cells from surgical specimens is presented. Tumor and normal tissues were processed by using the same methodology, based on mechanical disaggregation of tissue followed by enzymatic digestion and cell purification by sequential sieving. The overall procedure takes roughly one hour. The resulting cell preparations have excellent viabilities and yield. Establishment of primary cultures from all specimens was achieved successfully. The origin of primary cultured cells was established through morphological evaluation. Normal cells purity was confirmed by immunofluorescent staining and reverse transcription-polymerase chain reaction analysis for expression of specific markers.
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spelling pubmed-30877602011-05-13 A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens Valente, Maria João Henrique, Rui Costa, Vera L. Jerónimo, Carmen Carvalho, Félix Bastos, Maria L. de Pinho, Paula Guedes Carvalho, Márcia PLoS One Research Article The kidney is a target organ for the toxicity of several xenobiotics and is also highly susceptible to the development of malignant tumors. In both cases, in vitro studies provide insight to cellular damage, and represent adequate models to study either the mechanisms underlying the toxic effects of several nephrotoxicants or therapeutic approaches in renal cancer. The development of efficient methods for the establishment of human normal and tumor renal cell models is hence crucial. In this study, a technically simple and rapid protocol for the isolation and culture of human proximal tubular epithelial cells and human renal tumor cells from surgical specimens is presented. Tumor and normal tissues were processed by using the same methodology, based on mechanical disaggregation of tissue followed by enzymatic digestion and cell purification by sequential sieving. The overall procedure takes roughly one hour. The resulting cell preparations have excellent viabilities and yield. Establishment of primary cultures from all specimens was achieved successfully. The origin of primary cultured cells was established through morphological evaluation. Normal cells purity was confirmed by immunofluorescent staining and reverse transcription-polymerase chain reaction analysis for expression of specific markers. Public Library of Science 2011-05-04 /pmc/articles/PMC3087760/ /pubmed/21573239 http://dx.doi.org/10.1371/journal.pone.0019337 Text en Valente et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Valente, Maria João
Henrique, Rui
Costa, Vera L.
Jerónimo, Carmen
Carvalho, Félix
Bastos, Maria L.
de Pinho, Paula Guedes
Carvalho, Márcia
A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens
title A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens
title_full A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens
title_fullStr A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens
title_full_unstemmed A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens
title_short A Rapid and Simple Procedure for the Establishment of Human Normal and Cancer Renal Primary Cell Cultures from Surgical Specimens
title_sort rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3087760/
https://www.ncbi.nlm.nih.gov/pubmed/21573239
http://dx.doi.org/10.1371/journal.pone.0019337
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