Amastigote Culture
Amastigote Culture - Related Videos
04:46
Assessment of Leishmania Virulence within Cultured Macrophages
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2025
This video investigates Leishmania's virulence by treating macrophages with metacyclic promastigotes, resulting in the formation of intracellular amastigotes. The confirmation of Leishmania's virulence is achieved through nucleic acid fluorescent dye staining. This insight can contribute to the development of new therapies for leishmaniasis.
06:13
Introducing a Gene Knockout Directly Into the Amastigote Stage of Trypanosoma cruzi Using the CRISPR/Cas9 System
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Cited by 5 •
2019
Here, we describe a protocol to introduce a gene knockout into the extracellular amastigote of Trypanosoma cruzi, using the CRISPR/Cas9 system. The growth phenotype can be followed up either by cell counting of axenic amastigote culture or by proliferation of intracellular amastigotes after host cell invasion.
05:55
In Vivo Imaging of Leishmania infantum-infected Hamsters by Gingival Inoculation of Axenic Amastigotes Expressing Luciferase
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2025
Here we present a longitudinal evaluation of golden hamsters infected intraperitoneally (IP) or via intragingival (IG) with L. infantum-Luc by bioluminescent imaging and by PCR. Hamsters were evaluated 1 day post infection (1 dpi), 1 week after infection (8 dpi), and 3 weeks after infection (22 dpi) and euthanized at the 50th dpi and 8 months post infection.
08:17
Quantitative 3D Imaging of Trypanosoma cruzi-Infected Cells, Dormant Amastigotes, and T Cells in Intact Clarified Organs
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Cited by 3 •
2022
The present protocol describes light-sheet fluorescent microscopy and automated software-assisted methods to visualize and precisely quantify proliferating and dormant Trypanosoma cruzi parasites and T cells in intact, cleared organs and tissues. These techniques provide a reliable way to evaluate treatment outcomes and offer new insights into parasite-host interactions.
12:22
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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Cited by 118 •
2012
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.
FAQs
In axenic systems, researchers use increased temperature and acidic pH as environmental cues to induce promastigotes toward an amastigote-like state. These conditions model key changes associated with the parasite’s intracellular setting without requiring host cells. The resulting cultures allow investigators to examine stage-associated biology and gene-expression changes under controlled laboratory conditions.
Axenic cultures generate amastigote-like forms outside host cells, whereas macrophage infection models support parasite development within host cells. The axenic approach provides tighter control over experimental conditions, while the infection model places the parasite in a cellular environment relevant to intracellular survival. Comparing both systems can help separate intrinsic parasite responses from host-cell influences.
Stage-specific gene expression helps researchers determine how parasite genes behave as cells transition from promastigotes toward amastigote-like forms or develop inside macrophages. Changes in expression can be examined alongside virulence traits and intracellular survival. This connection is especially useful in genetics because it links a gene’s regulation with the parasite stage in which its function becomes relevant.
A study can begin with promastigotes and apply increased temperature and acidic pH to induce differentiation in an axenic system. Alternatively, researchers can use host-cell infection models in which macrophages support intracellular development. The selected route should match the question: controlled axenic conditions suit stage-focused comparisons, whereas macrophage models address development in a host-cell context.
Cultures provide material for testing how genetic changes affect stage-specific gene expression, virulence traits, and intracellular survival. Researchers can compare parasite lines or mutant phenotypes under axenic differentiation conditions or during macrophage infection. These comparisons help connect particular genetic alterations with altered developmental behavior or responses to experimental treatments.
The systems can reveal whether genetic differences are associated with altered differentiation, stage-specific expression, virulence-related traits, or intracellular survival. They also support examination of parasite responses to experimental treatments. Results from axenic and macrophage-based cultures can be interpreted together to determine whether an observed phenotype appears under controlled conditions, within host cells, or in both settings.