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Updated: Sep 11, 2025
Flow Cytometric Characterization of Murine B Cell Development
Published on: January 22, 2021
Assessment of flow cytometric tools to characterize milk somatic cells in water buffalo
Maria Carmela Scatà1, Francesco Grandoni1, Giovanna De Matteis1
1Council for Agricultural Research and Economics, CREA Research Centre for Animal Production and Aquaculture, Monterotondo (RM), Italy.
Insights
Researchers developed new flow cytometry methods to identify and characterize live milk somatic cells in water buffalo. This live differential cell count (LDCC) can aid in detecting mammary gland inflammation and monitoring milk quality.
Area of Science:
- Veterinary Immunology
- Dairy Science
- Flow Cytometry
Background:
- Accurate characterization of milk somatic cells is crucial for dairy animal health and milk quality assessment.
- Existing methods for somatic cell analysis in water buffalo milk are limited in their ability to differentiate specific cell types.
- Flow cytometry offers a potential high-throughput solution for detailed somatic cell analysis.
Purpose of the Study:
- To develop novel multicolour flow cytometric panels for the precise identification and characterization of live milk somatic cells in water buffalo (Bubalus bubalis).
- To establish a live differential cell count (LDCC) method for buffalo milk.
- To provide tools for future research into mammary gland health and milk technological properties.
Main Methods:
- Design and validation of four distinct multicolour antibody panels for flow cytometry.
- Utilized specific markers including CD18, CD172a, CD14, CD16, CD4, CD8, δ chain, CD335, CD79a, CD21, MHC-I, and pan Cytokeratin.
- Incorporated a Live/Dead vitality dye for distinguishing between live and dead cells in bulk milk samples.
Main Results:
- Panel 1 identified lymphocytes, polymorphonuclear neutrophils (PMN), and monocyte/macrophages.
- Panel 2 differentiated T helper (CD4+), T cytotoxic (CD8+), γδ T, and NK cells.
- Panel 3 characterized total and CD21+ B lymphocytes, while Panel 4 distinguished epithelial cells from leukocytes.
Conclusions:
- A robust flow cytometry-based method for live differential cell count (LDCC) in water buffalo milk has been established.
- The developed LDCC method enables fine characterization of live milk somatic cells.
- Future applications include identifying early inflammatory markers and assessing milk technological properties based on somatic cell composition.
Abstract:
The aim of this Research Communication was to develop new flow cytometric tools for the fine identification and characterization of milk somatic cells in water buffalo (Bubalus bubalis). Four multicolour panels of antibodies were designed to identify different subsets of live leukocytes and epithelial cells in bulk milk samples. Panel 1, including the CD18/CD172a/CD14/CD16 markers and Live/Dead vitality dye, allowed us to identify total lymphocytes, polymorphonuclear neutrophils (PMN) and monocyte/macrophage subsets. Panel 2 (CD18/CD4/CD8/δ chain/CD335 and Live/Dead dye) allowed us to identify T helper (CD4+), T cytotoxic (CD8+), γδ lymphocytes and NK cells. Panel 3 (CD18/CD79a/CD21 and Live/Dead dye) allowed us to identify total and CD21+ B lymphocytes. Finally, with Panel 4 (CD18/MHC-I/pan Cytokeratin and Live/Dead dye) the epithelial cells were distinguished from leukocytes. In conclusion, we propose a fine characterization of live milk somatic cell (live differential cell count (LDCC)) in buffalo species. In the future the determination of LDCC could used to identify new markers for detecting early inflammatory states of the mammary gland or for monitoring the technological properties of milks of different somatic cell composition.
