A technique for the isolation and purification of viable mucosal mast cells/globule leukocytes from the small intestine of parasitised sheep

• Published in: International journal for parasitology Vol. 24; no. 2; pp. 307 - 309
• Main Authors: Stankiewicz, M., Shaw, R.J., Jonas, W.E., Cabaj, W., Grimmett, D.J., Douch, P.G.C.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.04.1994; Elsevier Science
• Subjects: Animals; Biological and medical sciences; Cell Separation; Centrifugation, Density Gradient; eosinophils; globule leukocytes; Helminthic diseases; in vitro; Infectious diseases; Intestinal Mucosa - pathology; Intestine, Small - pathology; Leukocytes; Mast Cells; Medical sciences; Miscellaneous; Parasitic diseases; Sheep; Sheep Diseases - immunology; Trichostrongylosis - immunology; Trichostrongylosis - veterinary; Trichostrongylus colubriformis; globule leukocytes; eosinophils; mast cells; Trichostrongylus colubriformis; sheep; in vitro; Immunoprotection; Cell culture; Digestive system; Leukocyte; Gut; Parasite; Eosinophil; Vertebrata; Mammalia; Helmintha; Sheep; Nemathelminthia; Mast cell; Artiodactyla; Invertebrata; Nematoda; Viability; Ungulata
• ISSN: 0020-7519; 1879-0135
• DOI: 10.1016/0020-7519(94)90045-0

A technique for the isolation and purification of viable mucosal mast cells/globule leukocytes from the small intestine of parasitised sheep. Internationaljournal for Parasitology24: 307–309. Romney sheep, 1–2 years old, immunized by at least three anthelmintic abbreviated infections of 80–100,000 Trichostrongylus colubriformis larvae usually produced high numbers of intestinal mucosal mast cells/globule leukocytes (MMC/GLs). In isolating these cells, the importance of maintaining the intestine at 37°C, removal of mucus with dithiothreitol, enzymatic dispersion and careful in vitro handling procedures for maximising cell viability are emphasised. The MMC/GLs were separated from most contaminant cells by using a Percoll discontinuous gradient. MMC/GLs collected at the |60/100 % Percoll interface were passed through a complement coated nylon wool column to remove the contaminating eosinophils. Viable MMC/GLs were able to grow in vitro in the presence of Concanavalin A and survive in culture for up to 30 days. The MMC/ GLs were readily identified by ultraviolet light microscopy after staining with auramine O.