Lineage Depletion
Lineage Depletion - Related Videos
06:17
Depletion of Specific Cell Populations by Complement Depletion
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Cited by 15 •
2010
To effectively study the function of immune cell populations their purification is often required. Complement depletion is a fast and inexpensive technique for the isolation of immune cell populations with high purity.
08:50
Depletion and Reconstitution of Macrophages in Mice
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Cited by 118 •
2012
Macrophages play a central role in homeostasis and pathology in many tissues. The protocol presented here describes methods for depleting macrophages in vivo, deriving polarized macrophages from bone marrow aspirates, and adoptively transferring macrophages into mice. These techniques allow determination of the role that polarized macrophages play in health and disease.
11:07
Macrophage Cholesterol Depletion and Its Effect on the Phagocytosis of Cryptococcus neoformans
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Cited by 22 •
2014
In this article, a protocol for infection of macrophages with Cryptococcus neoformans is described. Also, a method for sterol depletion from the macrophages is explained. These protocols provide a guide to study fungal infections in vitro and examine the role of sterols in such infections.
02:45
Culturing Neisseria gonorrhoeae in Metal-Depleted Conditions
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2025
Source: Maurakis, S., and Cornelissen, C. N. Metal-Limited Growth of Neisseria gonorrhoeae for Characterization of Metal-Responsive Genes and Metal Acquisition from Host Ligands. J. Vis. Exp. (2020)This video demonstrates the procedure for growing Neisseria gonorrhoeae under metal-limited conditions using a defined, metal-depleted medium.
01:21
Lineage Commitment
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2023
Commitment is the process whereby stem cells: lose their ability to form all cell types and irreversibly change into a specific type. The multipotent hematopoietic stem cells, (HSCs), differentiate into the multipotent hematopoietic progenitor cells, (HPCs). The HPCs express many lineage-specific cytokine receptors. Each of these receptors binds specific cytokines, activates distinct signaling pathways, and expresses a particular gene set. The HPCs further differentiate to form committed...
FAQs
The antibody panel determines which cells are recognized as unwanted because each antibody targets a surface antigen associated with a mature immune, myeloid, erythroid, or other differentiated lineage. A panel that reflects the sample’s expected contaminants can remove those populations more effectively, leaving a fraction enriched for cells that lack the selected lineage markers.
The remaining fraction represents cells that do not express the mature lineage markers targeted by the antibody panel. It may contain less-differentiated populations, including hematopoietic stem and progenitor cells, but it is not necessarily a uniform population. Its composition depends on which markers were selected and which cells were removed during separation.
Removing abundant mature cells reduces background from contaminating populations, making less frequent cells easier to resolve in downstream experiments. This enrichment can improve the consistency of flow-cytometric measurements and help researchers examine stem or progenitor populations with less interference. The same principle supports clearer interpretation in differentiation assays, transplantation studies, and single-cell analysis.
Outcome depends on the relationship between the antibody panel and the surface markers present on unwanted cells. The sample’s mixture of mature immune, myeloid, erythroid, or other differentiated populations therefore matters, as does the effectiveness of labeling and subsequent capture or removal. These factors determine how completely contaminants are reduced and how useful the remaining fraction becomes.
A typical workflow begins by exposing the sample to antibodies that recognize surface antigens on unwanted mature cells. Those labeled cells are then captured or removed with magnetic beads or a related separation system. Researchers collect the remaining fraction for downstream analysis or experiments, such as flow cytometry, transplantation studies, differentiation assays, or single-cell analysis.
Researchers may choose this approach when mature lineage-positive cells would obscure a less-differentiated or rare population of interest. Reducing those cells before analysis can improve resolution and experimental consistency, particularly when the next step examines hematopoietic stem and progenitor cells. It is also useful when downstream assays require an enriched starting fraction rather than the original mixed sample.
In hematopoietic studies, depletion of mature immune, myeloid, erythroid, and other differentiated cells can enrich the sample for stem and progenitor populations. That enriched fraction can then be evaluated by flow cytometry, tested in transplantation studies, or used in differentiation assays. Single-cell analysis also benefits because reduced contamination can clarify the cellular populations detected.