Development and validation of a cell culture based assay for in vitro assessment of anticryptosporidial compounds
| Autor: | Arwid Daugschies, C. Wackwitz, Michael Najdrowski, Ute Mackenstedt, Damaris Kliemt, A. R. Heckeroth, Sandra Gawlowska |
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| Rok vydání: | 2007 |
| Předmět: |
Cryptosporidium
Cell Line Microbiology Apicomplexa chemistry.chemical_compound Piperidines medicine Animals Humans Parasite hosting Monensin Quinazolinones Dose-Response Relationship Drug General Veterinary Halofuginone biology General Medicine biology.organism_classification In vitro Infectious Diseases Cryptosporidium parvum chemistry Cell culture Insect Science Coccidiostats Parasitology Growth inhibition medicine.drug |
| Zdroj: | Parasitology Research. 101:161-167 |
| ISSN: | 1432-1955 0932-0113 |
| Popis: | In vitro culture of Cryptosporidium parvum oocysts in HCT-8 cells was combined with immunofluorescent labelling and digital image analysis to quantify the development of the parasite by detecting and measuring the labelled area in the respective cell cultures. The number of inoculated oocysts and the labelled area correlated reliably and significantly (R (2), 0.98-0.99). The effects of various concentrations of halofuginone bromide (0.00039 to 50 microM) and monensin (0.00225 to 0.144 microM) on in vitro parasite development were determined in further trials in cultures inoculated each with 10(5) oocysts. Monensin reduced the detected area in a dose-dependant manner. In comparison to the untreated controls, the area positive for C. parvum in the cultures treated with 0.144 to 0.009 microM monensin reached a maximum of 17%, and inhibition of 40% was observed at 0.0045 microM. Halofuginone bromide also efficiently inhibited parasite in vitro reproduction, albeit at higher concentrations. At 12.5 microM or more, inhibition was at least 90%; 0.05 microM still yielded 80% inhibition, whereas at concentrations below 0.00625 microM, labelled areas abruptly increased. Both drugs appeared efficient under in vitro conditions; the applied system is suited to screen drugs for their anti-cryptosporidial capacity. |
| Databáze: | OpenAIRE |
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