Novel method for continuous cell separation by density gradient centrifugation: evaluation of a miniature separation column.
A compact bench-top model of the centrifuge enables continuous cell separation based on density differences. The apparatus holds a small separation disk equipped with a circular channel (8 mL capacity) separated by a septum. A set of isotonic Percoll media with different densities is continuously introduced at one terminal and collected from the other. Under a centrifugal force field, cell suspension introduced into the proximal portion of the channel results in continuous separation of cells according to their densities. The performance of the apparatus was demonstrated with the separation of human buffy coat containing nucleated cells (>10(8)) among a large population (10(10)) of RBC. The results indicated that the method is capable of separating a large number of nucleated cells, with minimum damage, for a few hours of operation wherein neutrophils are well resolved from lymphocytes. The method may be applied to other types of samples including cord blood, blood from small animals, cultured cells, pancreatic beta cell islets, malaria parasites, sperm cells, etc.
Shiono
Hiroyuki
H
Laboratory of Biophysical Chemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-8014, USA.
Ito
Yoichiro
Y
D001772:Blood Cell Count / Q000379:methods; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D004912:Erythrocytes / Q000166:cytology; D006801:Humans; D007962:Leukocytes / Q000166:cytology; D008214:Lymphocytes / Q000166:cytology; D009000:Monocytes / Q000166:cytology; D009504:Neutrophils / Q000166:cytology; D011205:Povidone / Q000737:chemistry; D015203:Reproducibility of Results; D012822:Silicon Dioxide / Q000737:chemistry
C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone