HK-2: An immortalized proximal tubule epithelial cell line from normal adult human kidney
العنوان: | HK-2: An immortalized proximal tubule epithelial cell line from normal adult human kidney |
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المؤلفون: | J Kirk, Michael J. Ryan, Richard A. Zager, Sally M. Fuerstenberg, Gretchen Johnson, Beverly Torok-Storb |
المصدر: | Kidney International. 45:48-57 |
بيانات النشر: | Elsevier BV, 1994. |
سنة النشر: | 1994 |
مصطلحات موضوعية: | Adult, Male, medicine.medical_specialty, Renal cortex, Genetic Vectors, Molecular Sequence Data, Iron Chelating Agents, Kidney Tubules, Proximal, Reference Values, Epidermal growth factor, Internal medicine, medicine, Humans, Papillomaviridae, Cell Line, Transformed, Kidney, Base Sequence, biology, Histocytochemistry, Hydroxyl Radical, Cell growth, Acid phosphatase, Free Radical Scavengers, Hydrogen Peroxide, Immunohistochemistry, Molecular biology, Epithelium, medicine.anatomical_structure, Endocrinology, Nephrology, Cell culture, Molecular Probes, DNA, Viral, biology.protein, Alkaline phosphatase |
الوصف: | HK-2: An immortalized proximal tubule epithelial cell line from normal adult human kidney. Studies assessing mechanisms of proximal tubular cell (PTC) physiology and pathophysiology increasingly utilize cell culture systems to avoid the complexity of whole organ/whole animal experiments. However, no well-differentiated PTC line derived from adult human kidney currently exists. Therefore, the goal of this research was to establish such a line by transduction with human papilloma virus (HPV 16) E6/E7 genes. A primary PTC culture from normal adult human renal cortex was exposed to a recombinant retrovirus containing the HPV 16 E6/E7 genes, resulting in a cell line designated HK-2 (human kidney-2) which has grown continuously in serum free media for more than one year. HK-2 cell growth is epidermal growth factor dependent and the cells retain a phenotype indicative of well-differentiated PTCs (positive for alkaline phosphatase, gamma glutamyltranspeptidase, leucine aminopeptidase, acid phosphatase, cytokeratin, α 3 β 1 integrin, fibronectin; negative for factor VHI-related antigen, 6.19 antigen and CALLA endopeptidase). Furthermore, HK-2 cells retain functional characteristics of proximal tubular epithelium (Na + dependent/phlorizin sensitive sugar transport; adenylate cyclase responsiveness to parathyroid, but not to antidiuretic, hormone). The E6/E7 genes are present in the HK-2 genome, as determined by PCR. To assess its potential usefulness as a tool for studying injury and repair, HK-2 cells were exposed to a toxic concentration of H 2 O 2 ± iron chelation (deferoxamine) or hydroxyl radical scavenger (Na benzoate) therapy. Only the former blocked H 2 O 2 cytotoxicity, reproducing results previously obtained with freshly isolated rat proximal tubular segments. In conclusion, an immortalized adult human PTC line has been established by transduction with HPV 16 E6/E7 genes. It appears to be well-differentiated on the basis of its histochemical, immune cytochemical, and functional characteristics, and it can reproduce experimental results obtained with freshly isolated PTCs. Thus, this new PTC line could have substantial research application. |
تدمد: | 0085-2538 |
DOI: | 10.1038/ki.1994.6 |
URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4efa010b555c64b42740cdfbfa10578a https://doi.org/10.1038/ki.1994.6 |
Rights: | OPEN |
رقم الانضمام: | edsair.doi.dedup.....4efa010b555c64b42740cdfbfa10578a |
قاعدة البيانات: | OpenAIRE |
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Studies assessing mechanisms of proximal tubular cell (PTC) physiology and pathophysiology increasingly utilize cell culture systems to avoid the complexity of whole organ/whole animal experiments. However, no well-differentiated PTC line derived from adult human kidney currently exists. Therefore, the goal of this research was to establish such a line by transduction with human papilloma virus (HPV 16) E6/E7 genes. A primary PTC culture from normal adult human renal cortex was exposed to a recombinant retrovirus containing the HPV 16 E6/E7 genes, resulting in a cell line designated HK-2 (human kidney-2) which has grown continuously in serum free media for more than one year. 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