Collection and cultivation in vitro of equine mammary macrophages.

This research paper is about a study that successfully cultivated equine macrophages in a laboratory setting from female Shetland ponies using various culture mediums.

Methodology

• Macrophages were collected from the mammary gland of female Shetland ponies by injecting Escherichia coli lipopolysaccharide into the lactiferous ducts.
• After 6 to 11 days, a balanced salt solution was injected into the mammary gland to flush out accumulated cells.
• The mixture of harvested cells contained macrophages, lymphocytes, and neutrophils, with most of the cells being of the mononuclear type.

In-Culture Process

• Once in culture, the cells that adhered after 24 hours were identified as macrophages based on their morphological features, nonspecific esterase staining, and the presence of complement and immunoglobulin receptors.
• The macrophages were then cultivated in various media.
• A basal medium was found to be most effective in maintaining the spreading and adhesion of cells. This medium consisted of 15% equine serum, 10% bovine fetal serum, and RPMI 1640 medium containing 20 mM HEPES buffer.
• The researchers added a conditioned medium from mouse fibroblast cultures (L cells) at 30% to the basal medium. This improved the cell monolayers by reducing the formation of giant cells and also prolonged the adhesion of the cells.

Conclusions

• This methodology provides a new way of cultivating equine macrophages in vitro.
• The addition of a conditioned medium from mouse fibroblast cultures improves the viability of the macrophage cultures.