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

• Type: Article in International Scientific Journal
• Date: 2011

Authors

Maria Joao Valente (Author) Other; Rui Henrique (Author) ICBAS; Vera L Costa (Author) Other; Carmen Jeronimo (Author) ICBAS; Felix Carvalho (Author) FFUP; Maria L Bastos (Author) FFUP; de Pinho, PG (Author) FFUP; Marcia Carvalho (Author) Other

Journal

Title: PLoS ONE

Vol. 6 No. 8

Final page: e19337

ISSN: 1932-6203

Publisher: Public Library of Science

Indexing

ISI Web of Knowledge - 40 Citations

Scopus - 48 Citations

Other information

Authenticus ID: P-002-RJ5

DOI: 10.1371/journal.pone.0019337

Abstract (EN): 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.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8