Technical note: immunomagnetic procedure for positive selection of macrophages in ovine milk

Journal of Dairy Science
M CaropreseA Sevi

Abstract

A simple immunomagnetic procedure was developed to select macrophages from ovine milk by using a non-specific magnetic positive separation technique. Samples of ewe bulk milk were collected during early, mid, and late lactation; milk samples were centrifuged at 2,000 x g for 30 min at 4 degrees C; the fatty fraction and supernatant were removed, and each pellet was dissolved in 500 microL of pH 7.4 phosphate buffered saline + 0.02% NaN(3). Cells were targeted for selection by using mouse-IgG anti-ovine macrophages. Several trials, testing 2 different fluorochrome-conjugated antibodies [i.e., mouse anti-human CD14:R-Phycoerythrin (RPE) (MCA1568PE, Serotec) and F(ab')2 rabbit anti-mouse IgG:RPE (STAR12A, Serotec)] and 3 different labeling procedures, were performed to evaluate the purity of samples by flow cytometry. A morphological test was carried out by direct microscopic count in enriched fraction smears stained with May-Grünwald-Giemsa stain to confirm the presence of macrophages. The method described in the present technical note can be considered an innovative application to obtain a single-cell population of high purity selected from all the somatic cells in milk.

References

Jul 1, 1991·Journal of Dairy Science·R J Verdi, D M Barbano
May 23, 2007·Journal of Dairy Science·M CaropreseM Albenzio

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Citations

Dec 8, 2010·The Journal of Dairy Research·Marzia Albenzio, Mariangela Caroprese
Aug 9, 2018·The Journal of Dairy Research·Sara DivariFrancesca Tiziana Cannizzo
Dec 26, 2008·Journal of Dairy Science·M AlbenzioA Sevi
Mar 17, 2018·Biochemistry. Biokhimii︠a︡·A A Gordeev, A B Chetverin

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