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Mouse blood monocytes: standardizing their identification and analysis using CD115.

W L Breslin1, K Strohacker, K C Carpenter

  • 1Laboratory of Integrated Physiology, University of Houston, Houston, TX, United States.

Journal of Immunological Methods
|April 7, 2011
PubMed
Summary
This summary is machine-generated.

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Properly processing mouse blood monocytes is crucial for accurate immune studies. Refrigerating samples with EDTA and analyzing within 2 hours ensures reliable CD115 expression for flow cytometry analysis.

Area of Science:

  • Immunology
  • Hematology
  • Animal Models

Background:

  • Monocytes are key indicators of immune status in disease research.
  • Mouse models offer a valuable analog for studying immune responses longitudinally.
  • Standardized methods for analyzing mouse monocytes, particularly from small blood volumes, are lacking.

Purpose of the Study:

  • To establish optimal protocols for sample processing and flow cytometric analysis of mouse blood monocytes.
  • To identify reliable markers for mouse monocyte identification and enumeration.
  • To determine the stability of monocyte markers under various storage and processing conditions.

Main Methods:

  • Non-lethal blood collection from CD-1 male mice.
  • Comparison of monocyte markers (CD115, CD11b, CD14) for population identification.

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  • Assessment of CD115 expression stability at different time points (0, 2, 4h) and conditions (anticoagulant, temperature).
  • Evaluation of monocyte concentration consistency across varying blood volumes (5-40μL).

    • Main Results:

    • CD115, CD11b, and CD14 are effective markers for identifying mouse monocytes, with high co-expression rates observed.
    • CD115 expression significantly decreased after 4 hours at room temperature but remained stable when blood was treated with EDTA and refrigerated.
    • Accurate monocyte concentrations (<10% variation) were obtained using blood volumes as low as 10μL.

    Conclusions:

    • CD115 is a reliable marker for mouse monocyte identification in flow cytometry.
    • For delayed sample processing (>2h), treating blood with EDTA and refrigeration is essential to maintain CD115 stability.
    • These recommendations facilitate consistent and accurate analysis of mouse monocytes from small, non-lethal blood samples.