Author(s)

Ayumi Okamoto, Kosuke Torigata, Minami A. Sakurai, Daisuke Okuzaki, Hodaka Fujii, Toshinari Ohmine, Daisaku Miura, Shoichi Kimura, Norikazu Yabuta, Hiroshi Nojima

Combined Program on Microbiology and Immunology, Research Institute for Microbial Diseases.
Department of Molecular Genetics.
Department of Pharmacy, School of Pharmacy, Hyogo University of Health Sciences, 1-3-6 Minatoshima, Kobe, Hyogo 650-8530.
DNA-chip Development Center for Infectious Diseases.
Fukae Kasei Co., Ltd., 3-2-44 Takatsuka-dai, Nishi-ku, Kobe, Hyogo 651-2271, Japan.
Laboratory for Clinical Investigation, Osaka University Hospital; Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871.

Leukocytes from peripheral blood (PB) are of great value for diagnosis as well as basic and clinical research. However, no easy, centrifugation-free method is available for the isolation of live leukocytes from blood. We here develop a simple and quick method for the purification of viable leukocytes from whole blood using novel tools, named tLeukoCatch (tip-type) or sLeukoCatch (syringe-type), which is equipped with three Pall filter layers and captures leukocytes but not red blood cells (RBCs) in whole blood. Indeed, we showed that several million leukocytes per mL (~35% of the recovery rate) were captured and eluted from whole blood. The number of contaminant RBCs decreased from several million to several thousand. When mouse blood was hemolysed, almost all of the lysed RBC fragments were removed by passage through sLeukoCatch. Optical microscopic observation confirmed that the recovered leukocytes were sufficiently healthy to respond to growth stimuli. Efficient leukocyte recovery was also confirmed for hemolysed human blood. These results suggest that the LeukoCatch^TM system is useful for bedside diagnosis and basic research with blood samples.

Novel Tool; Leukocyte Purification; Centrifugation-Free; Diagnosis; PBMC

Okamoto, A. , Torigata, K. , Sakurai, M. , Okuzaki, D. , Fujii, H. , Ohmine, T. , Miura, D. , Kimura, S. , Yabuta, N. and Nojima, H. (2012) A simple and efficient method for the preparation of live leukocytes from peripheral blood using the LeukoCatch^TM system. Advances in Bioscience and Biotechnology, 3, 630-642. doi: 10.4236/abb.2012.35082.