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Updated: Dec 29, 2025
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Collection, Isolation, and Flow Cytometric Analysis of Human Endocervical Samples
Published on: July 6, 2014
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Optimization of processing female genital tissue samples for lymphocyte analysis by flow cytometry
Kevin A Stoner1, May A Beamer1, Hilary A Avolia1
1Magee-Womens Research and Foundation, Pittsburgh, PA, USA.
American Journal of Reproductive Immunology (New York, N.Y. : 1989)
|January 29, 2020
Summary
Optimizing genital tract specimen processing for immune cell analysis is crucial. Peripheral blood mononuclear cells (PBMCs) can model tissue processing effects on cell receptors, aiding method development.
Area of Science:
- Immunology
- Cell Biology
- Gynecologic Oncology
Background:
- Flow cytometry is vital for analyzing immune cells in genital tract specimens.
- Standardized processing methods are needed for accurate immune cell evaluation.
Purpose of the Study:
- To optimize processing of cervical cytobrush and genital tissue for flow cytometry.
- To evaluate peripheral blood mononuclear cells (PBMCs) as a model for tissue processing.
Main Methods:
- Processing of cytobrushes, PBMCs, and genital tissues under varied conditions.
- Enzymatic digestion with different collagenase types, concentrations, and durations.
- Assessing cell yield, viability, and surface receptor expression (e.g., CD3, CD4, CD8, CCR6, CD69).
Main Results:
- Collagenase B or D cleaved CCR6 expression; CD3, CD4, CD8 remained stable.
- Collagenase B increased CD69 expression, indicating cell activation.
- Delayed cytobrush processing or vortexing did not significantly impact immune cell viability or quantity.
Conclusions:
- Cervical biopsies require specific collagenase B treatment for optimal cell recovery.
- Endometrial biopsies can be processed using collagenase D.
- PBMCs serve as a reliable model for optimizing genital tract immune cell processing methods.
