A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimens
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10284/8091 |
Resumo: | 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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A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimensAgedCell culture techniquesFemaleHumansKidneyKidney neoplasmsMaleMiddle agedTumor cellsCulturedThe 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 ScienceRepositório Institucional da Universidade Fernando PessoaValente, Maria JoãoHenrique, RuiCosta, Vera L.Jerónimo, CarmenCarvalho, FélixBastos, Maria L.Guedes de Pinho, PaulaCarvalho, Márcia2019-10-01T16:00:31Z2011-05-01T00:00:00Z2011-05-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10284/8091eng1932-620310.1371/journal.pone.0019337info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2022-09-06T02:07:28Zoai:bdigital.ufp.pt:10284/8091Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T15:44:59.136234Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
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 |
spellingShingle |
A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimens Valente, Maria João Aged Cell culture techniques Female Humans Kidney Kidney neoplasms Male Middle aged Tumor cells Cultured |
title_short |
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_sort |
A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimens |
author |
Valente, Maria João |
author_facet |
Valente, Maria João Henrique, Rui Costa, Vera L. Jerónimo, Carmen Carvalho, Félix Bastos, Maria L. Guedes de Pinho, Paula Carvalho, Márcia |
author_role |
author |
author2 |
Henrique, Rui Costa, Vera L. Jerónimo, Carmen Carvalho, Félix Bastos, Maria L. Guedes de Pinho, Paula Carvalho, Márcia |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Institucional da Universidade Fernando Pessoa |
dc.contributor.author.fl_str_mv |
Valente, Maria João Henrique, Rui Costa, Vera L. Jerónimo, Carmen Carvalho, Félix Bastos, Maria L. Guedes de Pinho, Paula Carvalho, Márcia |
dc.subject.por.fl_str_mv |
Aged Cell culture techniques Female Humans Kidney Kidney neoplasms Male Middle aged Tumor cells Cultured |
topic |
Aged Cell culture techniques Female Humans Kidney Kidney neoplasms Male Middle aged Tumor cells Cultured |
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. |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011-05-01T00:00:00Z 2011-05-01T00:00:00Z 2019-10-01T16:00:31Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10284/8091 |
url |
http://hdl.handle.net/10284/8091 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1932-6203 10.1371/journal.pone.0019337 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Public Library of Science |
publisher.none.fl_str_mv |
Public Library of Science |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799130315865718784 |