Animals were sampled annually with approximately 52 weeks between sampling dates for the same animal unless medical management necessitated schedule alterations. Animals were sedated with intramuscular ketamine and femoral vein blood was drawn into sodium heparin-coated collection tubes (BD Bioscience). Complete blood counts (CBC) with differentials were collected at time of draw and then blood transferred to Duke by courier. The following day, plasma and peripheral blood mononuclear cells (PBMC) were separated using System-Histopaque-1077 separation media (Sigma). PBMC were washed in RPMI-1640 (Sigma), immunophenotyped and the remaining cells cryopreserved in 90% fetal bovine serum 10% dimethyl sulfoxide (Sigma).
System histopaque 1077 separation media
Manufactured by Merck Group
System-Histopaque-1077 separation media is a sterile, endotoxin-tested solution designed for the isolation of mononuclear cells from human peripheral blood. It is a density gradient medium used to separate cells based on their density differences.
Automatically generated - may contain errors
2 protocols using system histopaque 1077 separation media
1
Longitudinal Blood Sampling and PBMC Isolation
2
Longitudinal Blood Sampling and PBMC Isolation
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Animals were sampled annually with approximately 52 weeks between sampling dates for the same animal unless medical management necessitated schedule alterations. Animals were sedated with intramuscular ketamine and femoral vein blood was drawn into sodium heparin-coated collection tubes (BD Bioscience). Complete blood counts (CBC) with differentials were collected at time of draw and then blood transferred to Duke by courier. The following day, plasma and peripheral blood mononuclear cells (PBMC) were separated using System-Histopaque-1077 separation media (Sigma). PBMC were washed in RPMI-1640 (Sigma), immunophenotyped and the remaining cells cryopreserved in 90% fetal bovine serum 10% dimethyl sulfoxide (Sigma).
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