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A Parasite Rescue and Transformation Assay for Antileishmanial Screening Against Intracellular Leishmania donovani Amastigotes in THP1 Human Acute Monocytic Leukemia Cell Line
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Chapters
- 00:05Title
- 02:44Seeding and Differentiation of THP1 Macrophages in a 96-well Plate and 16-chamber Glass Culture Slide
- 04:13Infection of the Transformed THP1 Cells with Leishmania donovani Promastigotes
- 05:43Treatment of Infected Macrophages with Test Drugs
- 06:43Staining, Imaging and Image Analysis for Quantification of Infection and Drug Treatment Effect
- 08:09Parasite Rescue & Transformation Assay: Controlled Lysis of Parasite-infected Macrophages
- 08:59Quantitative Analysis of Transformed Promastigotes
- 09:22Results: % Infectivity and Parasite Survival in the Presence or Absence of Drug
- 10:41Conclusion
A parasite-rescue and transformation assay with THP1 cells infected in vitro with Leishmania donovani has been optimized for anti-leishmanial drug screening. The assay involves differentiation of THP1 cells, infection with promastigotes, treatment with test drugs, controlled lysis of the infected macrophages, rescue of amastigotes, transformation to promastigotes and monitoring promastigote growth and proliferation with a fluorometric assay.
Tags
Parasite RescueTransformation AssayAntileishmanial ScreeningIntracellular Leishmania Donovani AmastigotesTHP1 Human Acute Monocytic Leukemia Cell LineLeishmaniasisNeglected DiseasesDeveloping CountriesRisk Of Contracting LeishmaniasisVisceral LeishmaniasisLimited Drug OptionsTherapeutic Doses ToxicityMultiple Drug ResistanceAnti-leishmanial Drug Discovery PipelineIn Vitro Screening ModelsPromastigotes AssayAxenic Amastigotes AssayMacrophage-amastigotes Model
