title: A rapid and simple procedure for the establishment of human normal and cancer renal primary cell cultures from surgical specimens.
abstract: 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.
authors.affiliation: REQUIMTE - Laboratório de Toxicologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, Porto, Portugal. mjoao.pcv@gmail.com
authors.forename: Maria João
authors.initials: MJ
authors.lastname: Valente
authors.forename: Rui
authors.initials: R
authors.lastname: Henrique
authors.forename: Vera L
authors.initials: VL
authors.lastname: Costa
authors.forename: Carmen
authors.initials: C
authors.lastname: Jerónimo
authors.forename: Félix
authors.initials: F
authors.lastname: Carvalho
authors.forename: Maria L
authors.initials: ML
authors.lastname: Bastos
authors.forename: Paula Guedes
authors.initials: PG
authors.lastname: de Pinho
authors.forename: Márcia
authors.initials: M
authors.lastname: Carvalho
mesh_terms: D000368:Aged; D018929:Cell Culture Techniques / Q000379:methods*; D005260:Female; D006801:Humans; D007668:Kidney / Q000166:cytology*; D007680:Kidney Neoplasms / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D014407:Tumor Cells, Cultured