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Updated: Aug 8, 2026

Isolation of Precursor B-cell Subsets from Umbilical Cord Blood
Published on: April 16, 2013
The CD24 antigen discriminates between pre-B and B cells in human bone marrow
C Duperray1, J M Boiron, C Boucheix
1Service Commun INSERM de cytofluorométrie, INSERM U291, Montpellier, France.
Insights
CD24 antibody labeling identified two distinct B cell populations in adult bone marrow: CD24+ mature B cells and CD24(2+) early B lineage cells. This finding aids in understanding B cell development and differentiation.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- Bone marrow (BM) contains various lymphoid cell populations crucial for immune function.
- Understanding B cell lineage development requires precise identification of distinct cell subsets.
- CD24 is a cell surface antigen expressed on B lymphocytes, but its precise role in distinguishing subsets needs further clarification.
Purpose of the Study:
- To investigate the expression of CD24 on lymphoid cells in adult bone marrow.
- To differentiate and characterize distinct B cell populations based on CD24 expression levels.
- To elucidate the developmental stage and phenotype of CD24-defined B cell subsets.
Main Methods:
- Flow cytometry analysis of bone marrow lymphoid cells from healthy donors.
- Antibody labeling with CD24 and other B cell lineage markers (CD10, CD19, CD20, CD21, CD34, CD37, HLA-DR, surface IgM, cytoplasmic mu-chains).
- Dual fluorescence analysis to assess co-expression of markers on defined populations.
Main Results:
- Two CD24-positive populations were identified: CD24+ (intermediate density) and CD24(2+) (high density).
- CD24+ cells exhibited a mature B cell phenotype (CD19+, CD21+, CD37+, sIgM+, HLA-DR+).
- CD24(2+) cells represented early B lineage cells (CD19+, HLA-DR+, CD34+, with varying expression of CD10, CD20, and cytoplasmic mu-chains).
Conclusions:
- CD24 antigen expression effectively discriminates between mature B cells and early B lineage precursors in adult bone marrow.
- The CD24(2+) population encompasses various pre-B cell stages, including "pre" pre-B, "intermediate" pre-B, and "true" pre-B cells.
- The CD24+ population corresponds to B cells with a standard mature phenotype.
Abstract:
When bone marrow (BM) lymphoid cells from 12 adult healthy donors were labeled by CD24 antibodies and analyzed by flow cytometry, two positive populations of cells were demonstrated in each sample (by a separated bimodal specific immunofluorescence). One population had intermediate CD24-Ag density (termed CD24+ cells) whereas the other had high CD24-Ag density (termed CD24(2+) cells). CD24+ cells represented 5.8 +/- 2.7% of the total lymphoid BM cells and CD24(2+) cells 5.6 +/- 2.5%. Using dual fluorescence analysis on eight samples, all CD24+ cells expressed the CD21 and CD37 mature B cell Ag and also surface IgM (sIgM), but this population lacked CD10 Ag. These cells also expressed CD19 Ag, and at a higher density than CD24(2+) cells. They were also positive for HLA-DR Ag. Conversely, CD24(2+) cells were shown to be early cells of the B cell lineage. While all the CD24(2+) cells were HLA-DR+ and CD19+, 64 +/- 16% of them expressed CD20 Ag (at a lower density than CD24+ cells), 65 +/- 21% CD10 Ag, and 22 +/- 8% were positive for cytoplasmic mu-chains (c mu). None of these cells expressed the CD21 and CD37 mature B cell Ag or sIgM. Additional experiments on four different healthy donors demonstrated that 30 +/- 9% of the CD24(2+) cells expressed the CD34 Ag and that the CD24+ cells did not express it. Thus, the CD24 Ag permits discrimination between two populations of the B cell lineage present in adult BM: 1) A CD24(2+) cell population including "pre" pre-B cells (HLA-DR+, CD19+, CD10+/-, CD20-, CD21-, CD34+, CD37-, c mu-), "intermediate" pre-B cells (HLA-DR+, CD19+, CD10+, CD20+, CD21-, CD34-, CD37-, c mu-), and "true" pre-B cells (HLA-DR+, CD19+, CD10+, CD20+, CD21-, CD34-, CD37-, c mu+). 2) A CD24+ cell population including B cells of the standard phenotype (HLA-DR+, CD19+, CD10-, CD20+, CD21+, CD34-, CD37+, c mu-, sIgM+).
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