Isolation of basal guinea pig granulocytes for the study of pulmonary inflammation.
Siegel-PD; James-AW; Clark-PR; Castanova-V
Agricultural health and safety: workplace, environment, sustainability, supplement. McDuffie HH, Dosman JA, Semchuk KM, Olenchock SA, Senthilselvan A, eds. Saskatoon, Saskatchewan, Canada: University of Saskatchewan, 1994 Oct; :59-66
A technique for isolating basal (control) guinea-pig granulocytes for use in studies of pulmonary inflammation was developed. The purpose of the experiments was to develop an animal model suitable for examining the role of granulocytes in organic dust toxic syndrome and other pulmonary inflammatory disorders. Spleens were removed from naive English-short-haired-guinea-pigs, sieved through a 40 mesh tissue ultrasieve and then through a 330T mono nylon screen. The splenocytes were then pelleted by centrifugation for 5 minutes. The pellet was resuspended and centrifuged at 2,000 revolutions per minute using a centrifuge and elutriation rotor. The granulocyte enriched fraction was then elutriated. The granulocytes were again pelleted, resuspended, and counted using a Coulter counter. The purified granulocyte fraction produced by this procedure accounted for an average of 34.87% of the total splenocyte population. The average viability of the granulocytes, as determined by the trypan-blue dye exclusion test, was 83.6%. Addition of opsonized zymosan to the granulocyte preparation stimulated production of superoxide anion and hydrogen-peroxide. The lack of formation of superoxide and hydrogen-peroxide following treatment with unopsonized zymosan indicated that the granulocyte preparation was relatively uncontaminated by monocytes. The authors conclude that this procedure can isolate purified granulocyte fractions useful for studying the role of granulocytes in pulmonary inflammation.
Mammalian-cells; Laboratory-animals; Splenic-tissue; Cellular-function; Laboratory-techniques; In-vitro-studies; Bioassays; Respiratory-system-disorders
McDuffie-HH; Dosman-JA; Semchuk-KM; Olenchock-SA; Senthilselvan-A
Agricultural health and safety: workplace, environment, sustainability, supplement