The selection of serum-independent PC12 cells for a more-reliable manganese cytotoxicity test
| Autor: | Antonella Fusari, Antonio Ubaldi, Alessandra Gorreri, Giovanni Del Bono, Gaetano L. Aiello, Daniela Franchi, Enrico Bignetti |
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| Přispěvatelé: | BIGNETTI E, FRANCHI D, GORRERI A, FUSARI A, DEL BONO G, UBALDI A, AIELLO GL |
| Rok vydání: | 2007 |
| Předmět: |
Cell Survival
Adrenal Gland Neoplasms Pheochromocytoma Toxicology Animal Testing Alternatives PC12 Cells General Biochemistry Genetics and Molecular Biology Culture Media Serum-Free law.invention chemistry.chemical_compound law Doubling time Cytotoxic T cell Animals Cytotoxicity Manganese biology Chemistry Petri dish Resazurin General Medicine Enzyme assay In vitro Rats Medical Laboratory Technology Biochemistry Toxicity biology.protein |
| Zdroj: | Alternatives to laboratory animals : ATLA. 35(4) |
| ISSN: | 0261-1929 |
| Popis: | A major issue concerning the protocols of heavy metal cytotoxicity tests with PC12 cells was the hypothesis that serum in the culture medium might sequester the metal, thus altering the results obtained. However, serum withdrawal impairs the viability of PC12 cells themselves, thus impeding cytotoxicity testing in the absence of serum. In this study, we repeatedly selected undifferentiated, totally non-adherent PC12 cells in Petri dishes. Surprisingly, we discovered that these cells could survive and proliferate in serum-free medium. Moreover, features such as NGF-responsiveness, resazurin reduction potential, doubling rate, protein content, and basal caspase-3 enzyme activity, were equivalent to those exhibited by standard PC12 cultures. Further experiments aimed at fully characterising these serum-independent PC12 cells are in progress. These cells enabled cytotoxicity experiments to be conducted with manganese, both in serum-supplemented and in serum-deprived medium. The results demonstrated that serum removal decreased the LC50 of manganese from 250μM to 32μM, without affecting the internalisation of the metal. The data exclude an early competitive effect of serum on metal internalisation; rather, they suggest a late protective mechanism mediated by serum against the cytotoxic effect of the already-internalised metal. |
| Databáze: | OpenAIRE |
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