The Direct Lineage Cell Depletion Kit and the indirect Lineage Cell Depletion Kit have been developed for the efficient depletion of mature hematopoietic cells, including T cells, B cells, monocytes/macrophages, granulocytes, erythrocytes and their committed precursors. Both kits are optimized for untouched isolation of lineage-negative cells from mouse bone marrow samples. Using the direct isolation method, depletion can be automated and completed in 20 minutes.

Data and images for Lineage Cell Depletion Kit, mouse

Figures

Figure 1

Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant
®
Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
Before separation
After separation
LSK staining
View details

Figure 1

Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant
®
Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
View details

Figure 1

Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant
®
Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
View details

Figure 1

Isolation of lineage marker-negative cells from mouse bone marrow using the Direct Lineage Cell Depletion Kit, a MidiMACS™ Separator and an LS Column. Cells were fluorescently labeled with the Lineage Cell Detection Cocktail-Biotin and Anti-Biotin-APC and analyzed by flow cytometry using the MACSQuant
®
Analyzer. In order to evaluate the LSK fraction, cells were additionally labeled with CD117-PE and Anti-SCA-1-FITC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Specifications for Lineage Cell Depletion Kit, mouse

Overview

The Direct Lineage Cell Depletion Kit and the indirect Lineage Cell Depletion Kit have been developed for the efficient depletion of mature hematopoietic cells, including T cells, B cells, monocytes/macrophages, granulocytes, erythrocytes and their committed precursors. Both kits are optimized for untouched isolation of lineage-negative cells from mouse bone marrow samples. Using the direct isolation method, depletion can be automated and completed in 20 minutes.

Detailed product information

Detailed separation procedure

For depletion, cells are magnetically labeled with an antibody cocktail against so-called lineage markers (CD5, CD45R (B220), CD11b, Gr-1 (Ly-6G/C), 7-4, and Ter-119). In the case of the Lineage Cell Depletion Kit, mouse, the labeling is performed in a two-step procedure. Cells are first labeled with a cocktail of biotinylated antibodies followed by labeling with Anti-Biotin MicroBeads.
In the case of the Direct Lineage Cell Depletion Kit the antibodies are directly coupled to MicroBeads allowing a time-saving one-step labeling and isolation of lineage-negative cells in only 20 minutes.

Downstream applications

The Lineage Cell Depletion Kit has been used for the enrichment of both, hematopoietic stem cells (HSCs)
4
and marrow stromal cells (MSCs) from bone marrow
5
. The labeling procedure leaves lineage-negative cells untouched, allowing further magnetic separation of lineage-negative cells according to expression of markers such as CD117
1,2
or Sca-1
3
.

Columns

LS Columns, autoMACS
®
Columns, or Multi-24 Column Blocks.

Resources for Lineage Cell Depletion Kit, mouse

Documents and Protocols

Reviews for Lineage Cell Depletion Kit, mouse

Mouse Lineage Depletion Kit Review

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Lineage Cell Depletion Kit, mouse (130-090-858)

Our lab has generated a mouse model that expresses three transgenic human host factors and can support HIV infection. We are interested in understanding protective immune responses that could protect against HIV infection in these mice. To understand the role that CD8 T cell responses might have against HIV infection, we reprogram their precursor cells at stem cell level by introducing TCRs that are known to have potent anti-HIV activity. We use this kit to deplete all lineage committed cells from bone marrow.

References for Lineage Cell Depletion Kit, mouse

Publications

  1. Orlic, D. (2002) Stem cell repair in ischemic heart disease: an experimental model. Int. J. Hematol. 76(suppl.1): 144-145
  2. Orlic, D. et al. (2001) Mobilized bone marrow cells repair the infarcted heart, improving function and survival. Proc. Natl. Acad. Sci. U.S.A. 98: 10344-10349
  3. Lagasse, E. et al. (2000)
    Purified hematopoietic stem cells can differentiate into hepatocytes
    in vivo
    .
    Nat Med 6: 1229-1234
  4. Tang, H. et al. (2006) Endothelial stroma programs hematopoietic stem cells to differentiate into regulatory dendritic cells through IL-10. Blood 108: 1189-1197
  5. Forte, G. et al. (2006) Hepatocyte growth factor effects on mesenchymal stem cells: proliferation, migration, and differentiation. Stem Cells 24: 23-33

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