Inkt Cell Isolation
Inkt Cell Isolation - Related Videos
03:35
Isolating Invariant Natural Killer T Cells from a Mouse Model
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2025
This video illustrates a method for isolating invariant natural killer T cells, also known as iNKT cells. The procedure begins with injecting a synthetic glycolipid into a mouse to stimulate the activation and proliferation of iNKT cells in the spleen. Subsequently, single-cell suspensions are prepared from the spleen and incubated with iNKT-cell-recognizing complexes labeled with fluorophores, followed by magnetic bead-based separation, to isolate the iNKT cells.
09:01
An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen
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Cited by 16 •
2015
Here we present an adapted protocol that can be used to generate a large number of murine invariant natural killer T cells from mouse spleen. The protocol outlines an approach by which splenic iNKT cells can be enriched for, isolated and expanded in vitro using a limited number of animals and reagents.
08:43
Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice
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Cited by 2 •
2020
This protocol uses G6PI mixed peptides to construct rheumatoid arthritis models that are closer to that of human rheumatoid arthritis in CD4+ T cells and cytokines. High purity invariant natural killer T cells (mainly iNKT2) with specific phenotypes and functions were obtained by in vivo induction and in vitro purification for adoptive immunotherapy.
10:33
Isolation and Ex Vivo Culture of Vδ1+CD4+γδ T Cells, an Extrathymic αβT-cell Progenitor
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2015
Here, we provide an optimized protocol for the isolation and cloning of the scarce T-cell entity of peripheral Vδ1+CD4+ T cells that is, as we showed recently, an extrathymic αβ T-cell progenitor. This technique allows to quantitatively isolate, clone and efficiently expand these cells in ex vivo culture.
07:26
Isolation of Functional Cardiac Immune Cells
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Cited by 9 •
2011
This method for isolating functional immune cells from the heart provides an alternative to the conventional methods of collagenase digestion, which causes unwanted immune cell activation, resulting in a decreased responsiveness of these cells. Our method of isolation yields functional cardiac immune cells by avoiding problems associated with enzymatic digestion.
FAQs
Identification relies on the combined detection of the invariant T-cell receptor and natural killer cell markers rather than on a single feature alone. Flow cytometry can measure these markers in the separated population and, when needed, sort the identified cells for further study. This approach connects phenotypic confirmation with downstream analyses of iNKT-cell function.
Each step addresses a different part of the separation process. Sample preparation establishes a workable starting suspension, density-based separation helps partition blood or tissue components, and antibody-mediated magnetic enrichment concentrates the desired lymphocytes. Flow-cytometric identification or sorting then verifies the target population through marker expression, producing cells suitable for controlled immunological analysis.
Antibody-mediated magnetic enrichment is used to concentrate iNKT cells from a prepared sample, whereas flow cytometry identifies cells according to the invariant T-cell receptor and natural killer cell markers and can sort the selected population. The two methods can therefore operate sequentially: enrichment reduces the starting complexity, while cytometric analysis provides marker-based confirmation or further selection.
Once separated, iNKT cells can be examined for cytokine production, activation, and cytotoxicity, allowing investigators to connect cell identity with immune behavior. Their interactions with antigen-presenting cells can also be studied in controlled conditions. These readouts help characterize how iNKT cells participate in immune responses rather than only documenting their presence in blood or tissue.
A typical workflow begins with sample preparation, followed when appropriate by density-based separation of the blood or tissue material. Antibody-mediated magnetic enrichment can then concentrate the target lymphocytes. Flow-cytometric identification, and potentially sorting, uses the invariant T-cell receptor and natural killer cell markers to define the population for subsequent functional or interaction studies.
The approach is useful when investigators need to examine iNKT-cell responses to microbial infection or inflammatory conditions in a controlled system. Isolated cells can be evaluated for cytokine production, activation, cytotoxicity, or interactions with antigen-presenting cells. These measurements provide subject-specific context for studying how lymphocytes linking innate and adaptive immunity respond during disease-related processes.
In cancer immunology, separated iNKT cells can be analyzed for activation, cytotoxicity, cytokine production, and interactions with antigen-presenting cells. Those measurements help researchers investigate their potential role in tumor-related immune responses. The same isolation strategy also supports development of iNKT-directed diagnostics or therapies by providing a defined cell population for controlled immunological study.