A two-step immunomagnetic separation of somatic cell subpopulations for a gene expression profile study in bovine milk
| Autor: | Laura Starvaggi Cucuzza, Paola Pregel, Paola Sacchi, Francesca Tiziana Cannizzo, Fulvio Riondato, Roberto Rasero, Sara Divari, Bartolomeo Biolatti |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Somatic cell Neutrophils Cell Mastitis Biology Immunomagnetic separation Monocytes 03 medical and health sciences Gene expression medicine Animals Lymphocytes RNA Messenger magnetic separation Gene Mastitis Bovine innate immunity Innate immune system Immunomagnetic Separation milk somatic cells RNA General Medicine Staphylococcal Infections medicine.disease Molecular biology Immunity Innate 030104 developmental biology medicine.anatomical_structure Milk gene expression Cattle Female Animal Science and Zoology Transcriptome Food Science |
| Popis: | The objective of this study was to demonstrate the usefulness of an immunomagnetic method to purify subpopulations of milk somatic cells. The experiment was conducted on milk samples collected from healthy cows (n = 17) and from cows with clinical mastitis (n = 24) due to a Staphylococcus aureus natural infection. A two-step immunomagnetic purification was applied to simultaneously separate three somatic cell subpopulations from the same milk sample. Total RNA was extracted and qPCR was performed to determinate mRNA levels of innate immunity target genes in purified somatic cell subpopulations. Good quality and quantity of RNA allowed the reference gene analysis in each cell subpopulation. An up-regulation of the main genes involved in innate immune defence was detected in separated polymorphonuclear neutrophilic leucocytes-monocytes and lymphocytes of mastitic milk. These results and flow cytometric analysis suggest that the immunomagnetic purification is an efficient method for the isolation of the three populations from milk, allowing the cells to be studied separately. |
| Databáze: | OpenAIRE |
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