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Updated: Aug 8, 2026
Analysis of Cell Cycle Position in Mammalian Cells
Published on: January 21, 2012
Cell-cycle kinetics of proliferating mouse B lymphocytes in vitro
1Immunology Unit, Lund University, Sweden.
Insights
Different stimuli affect B lymphocyte proliferation rates. Anti-kappa antibodies induce faster cell-cycle entry and higher proliferation frequency in B lymphocytes compared to lipopolysaccharide, though growth kinetics remain constant.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- B lymphocytes are key players in adaptive immunity.
- Understanding B lymphocyte activation and proliferation is crucial for immunology research.
- In vitro stimulation models are essential for studying lymphocyte responses.
Purpose of the Study:
- To compare the cell-cycle kinetics of B lymphocytes stimulated with different agents.
- To determine the effect of lipopolysaccharide versus anti-kappa antibodies on B lymphocyte proliferation.
- To analyze the frequency and kinetics of B lymphocyte cell-cycle entry.
Main Methods:
- In vitro culture of resting mouse B lymphocytes.
- Stimulation using lipopolysaccharide and Sepharose-coupled anti-kappa antibodies.
- Cell cycle analysis via DNA staining and M phase block.
- Determination of labeling index and generation time.
Main Results:
- Anti-kappa antibody stimulation resulted in more rapid cell-cycle entry and higher frequency compared to lipopolysaccharide.
- The labeling index was 85% for anti-kappa stimulated cells versus 60% for lipopolysaccharide stimulated cells.
- The generation time for actively cycling B lymphocytes was approximately 18 hours, irrespective of the stimulus.
Conclusions:
- The fraction of B lymphocytes induced to proliferate in vitro is stimulus-dependent.
- The growth kinetics of proliferating B lymphocyte populations are consistent across different stimuli.
- This study highlights differential B lymphocyte activation pathways.
Abstract:
Resting mouse B lymphocytes were stimulated in vitro with lipopolysaccharide, Sepharose-coupled anti-kappa antibodies or a combination of the two. B lymphocytes stimulated with anti-kappa entered the cell-cycle with more rapid kinetics and at a higher frequency than did the corresponding cell population stimulated with lipopolysaccharide. Using cell cycle analysis after DNA staining combined with an M phase block, the cell-cycle kinetics of in vitro cultured B-lymphocytes was studied. The labelling index of lipopolysaccharide stimulated B lymphocytes was 60% while that for anti-kappa Sepharose stimulated cells was 85%. The generation time of the actively cycling population from both types of cultures was constant and was of the order of 18 h. Thus, the fraction of B lymphocytes induced to proliferate in vitro varies depending on the stimulus, while the growth kinetics of the actively proliferating populations are remarkably constant.
