Simple and cost-effective isolation of monocytes from buffy coats.
We have optimized the procedure of monocyte isolation on a Percoll density gradient. The new procedure consists of three steps: (1) the isolation of MNC on a Ficoll density gradient; (2) the separation of monocytes from lymphocytes on a high-density hyper-osmotic Percoll density gradient; and (3) the separation of monocytes from platelets and dead cells on a low-density iso-osmotic Percoll density gradient. The procedure is simple and cost-effective. Monocyte purity and recovery are both about 75% and platelet contamination is low. The isolated monocytes retain their capacity to differentiate into dendritic cells in vitro.
Repnik
Urska
U
Blood Transfusion Centre of Slovenia, Tissue Typing Centre, Slajmerjeva 6, 1000 Ljubljana, Slovenia. urska.repnik@guest.arnes.si
Knezevic
Miomir
M
Jeras
Matjaz
M
D002454:Cell Differentiation; D019070:Cell Lineage; D002469:Cell Separation / Q000191:economics / Q000379:methods*; D002470:Cell Survival; D002499:Centrifugation, Density Gradient / Q000379:methods*; D003362:Cost-Benefit Analysis; D005362:Ficoll; D006801:Humans; D009000:Monocytes / Q000166:cytology*; D011205:Povidone; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide
D005362:Ficoll; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone