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Cell culture system for studying bovine neutrophil diapedesis

(1996) JOURNAL OF DAIRY SCIENCE. 79(8). p.1353-1360
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Abstract
Neutrophils are the major defense against bacterial infection in the bovine mammary gland. Neutrophils migrate from blood into the lumen of the gland in response to inflammatory stimuli. This study describes the development of a system of cell culture that can be used to study neutrophil diapedesis through secretory and ductal mammary epithelial barriers. The culture system consists of successive layers of collagen, fibroblasts, collagen, and a confluent monolayer of secretory or ductal epithelial cells layered on a porous membrane. Confluence was determined by electrical resistance and trypan blue diffusion. Neutrophil diapedesis occurred from the basal to the apical surface of the monolayers. Purified complement C5a, fetal bovine serum that had been activated by zymosan, and fetal bovine serum that had been activated by Escherichia coli induced neutrophil diapedesis. Neutrophil diapedesis was greater across ductal cell monolayers. Blood neutrophils from five cows differed in their ability to migrate through the multilayered culture system in response to C5a. Monoclonal antibodies to C5a blocked diapedesis induced by purified C5a but had no effect on diapedesis induced by fetal bovine serum that had been activated by zymosan or by fetal bovine serum that had been activated by E. coli endotoxin, indicating that factors other than C5a were chemotactic for neutrophils. Monomeric IgG(2), immune complexes, and E. coli endotoxin did not induce neutrophil diapedesis.
Keywords
MIGRATION, CHEMOTAXIS, MAMMARY-GLAND, IMMUNE-COMPLEXES, STAPHYLOCOCCUS-AUREUS, POLYMORPHONUCLEAR LEUKOCYTES, IN-VITRO, cell culture, neutrophil, diapedesis, DEPENDENCE, COMPLEMENT, EPITHELIUM

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Citation

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Chicago
Smits, Elke, E Cifrian, AJ Guidry, P Rainard, Christian Burvenich, and Max Paape. 1996. “Cell Culture System for Studying Bovine Neutrophil Diapedesis.” Journal of Dairy Science 79 (8): 1353–1360.
APA
Smits, E., Cifrian, E., Guidry, A., Rainard, P., Burvenich, C., & Paape, M. (1996). Cell culture system for studying bovine neutrophil diapedesis. JOURNAL OF DAIRY SCIENCE, 79(8), 1353–1360.
Vancouver
1.
Smits E, Cifrian E, Guidry A, Rainard P, Burvenich C, Paape M. Cell culture system for studying bovine neutrophil diapedesis. JOURNAL OF DAIRY SCIENCE. 1996;79(8):1353–60.
MLA
Smits, Elke, E Cifrian, AJ Guidry, et al. “Cell Culture System for Studying Bovine Neutrophil Diapedesis.” JOURNAL OF DAIRY SCIENCE 79.8 (1996): 1353–1360. Print.
@article{189993,
  abstract     = {Neutrophils are the major defense against bacterial infection in the bovine mammary gland. Neutrophils migrate from blood into the lumen of the gland in response to inflammatory stimuli. This study describes the development of a system of cell culture that can be used to study neutrophil diapedesis through secretory and ductal mammary epithelial barriers. The culture system consists of successive layers of collagen, fibroblasts, collagen, and a confluent monolayer of secretory or ductal epithelial cells layered on a porous membrane. Confluence was determined by electrical resistance and trypan blue diffusion. Neutrophil diapedesis occurred from the basal to the apical surface of the monolayers. Purified complement C5a, fetal bovine serum that had been activated by zymosan, and fetal bovine serum that had been activated by Escherichia coli induced neutrophil diapedesis. Neutrophil diapedesis was greater across ductal cell monolayers. Blood neutrophils from five cows differed in their ability to migrate through the multilayered culture system in response to C5a. Monoclonal antibodies to C5a blocked diapedesis induced by purified C5a but had no effect on diapedesis induced by fetal bovine serum that had been activated by zymosan or by fetal bovine serum that had been activated by E. coli endotoxin, indicating that factors other than C5a were chemotactic for neutrophils. Monomeric IgG(2), immune complexes, and E. coli endotoxin did not induce neutrophil diapedesis.},
  author       = {Smits, Elke and Cifrian, E and Guidry, AJ and Rainard, P and Burvenich, Christian and Paape, Max},
  issn         = {0022-0302},
  journal      = {JOURNAL OF DAIRY SCIENCE},
  keyword      = {MIGRATION,CHEMOTAXIS,MAMMARY-GLAND,IMMUNE-COMPLEXES,STAPHYLOCOCCUS-AUREUS,POLYMORPHONUCLEAR LEUKOCYTES,IN-VITRO,cell culture,neutrophil,diapedesis,DEPENDENCE,COMPLEMENT,EPITHELIUM},
  language     = {eng},
  number       = {8},
  pages        = {1353--1360},
  title        = {Cell culture system for studying bovine neutrophil diapedesis},
  url          = {http://jds.fass.org/cgi/content/abstract/79/8/1353},
  volume       = {79},
  year         = {1996},
}