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Isolation of basal guinea pig granulocytes for the study of pulmonary inflammation.

Authors
Siegel-PD; James-AW; Clark-PR; Castanova-V
Source
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
Link
NIOSHTIC No.
00238788
Abstract
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.
Keywords
Mammalian-cells; Laboratory-animals; Splenic-tissue; Cellular-function; Laboratory-techniques; In-vitro-studies; Bioassays; Respiratory-system-disorders
Publication Date
19941001
Document Type
Conference/Symposia Proceedings
Editors
McDuffie-HH; Dosman-JA; Semchuk-KM; Olenchock-SA; Senthilselvan-A
Fiscal Year
1995
NTIS Accession No.
NTIS Price
NIOSH Division
DRDS
Priority Area
Respiratory-system-disorders
Source Name
Agricultural health and safety: workplace, environment, sustainability, supplement
State
WV
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