ABB  Vol.3 No.5 , September 2012
A simple and efficient method for the preparation of live leukocytes from peripheral blood using the LeukoCatchTM system
Abstract: 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 LeukoCatchTM system is useful for bedside diagnosis and basic research with blood samples.
Cite this paper: 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 LeukoCatchTM system. Advances in Bioscience and Biotechnology, 3, 630-642. doi: 10.4236/abb.2012.35082.

[1]   Chen DY, Hsieh TY, Hsieh CW, Lin FJ, Lan JL (2007) Increased apoptosis of peripheral blood lymphocytes and its association with interleukin-18 in patients with active untreated adult-onset Still's disease. Arthritis Rheum, 57, 1530-1538.

[2]   Kitamura T, Kabuyama Y, Kamataki A, Homma MK, Kobayashi H, et al. (2008) Enhancement of lymphocyte migration and cytokine production by ephrinB1 system in rheumatoid arthritis. Am J Physiol Cell Physiol, 294, 189-196.

[3]   Attur M, Belitskaya-Lévy I, Oh C, Krasnokutsky S, Greenberg J, et al. (2011) Increased interleukin-1β gene expression in peripheral blood leukocytes is associated with increased pain and predicts risk for progression of symptomatic knee osteoarthritis. Arthritis Rheum, 63, 1908-1917.

[4]   Mellman I, Coukos G, Dranoff G (2011) Cancer immunotherapy comes of age. Nature, 480, 480-489.

[5]   Curran KJ, Pegram HJ, Brentjens RJ (2012) Chimeric antigen receptors for T cell immunotherapy: current understanding and future direction. J Gene Med, 6, 405-415.

[6]   Yu M, Stott S, Toner M, Maheswaran S, Haber DA (2011) Circulating tumor cells: approaches to isolation and characterization. J Cell Biol, 192, 373-382.

[7]   Boyum A (1968) Isolation of mononuclear cells and granulocytes from human blood (Paper IV) . Scand J Clin Lab Invest Suppl, 97, 77–89.

[8]   B?yum A, Brincker Fjerdingstad H, Martinsen I, Lea T, L?vhaug D (2002) Separation of human lymphocytes from citrated blood by density gradient (NycoPrep) centrifugation: monocyte depletion depending upon activation of membrane potassium channels. Scand J Immunol, 56, 76-84.

[9]   Tomana M, Parton A, Barnes DW (2008) An improved method for separation of leucocytes from peripheral blood of the little skate (Leucoraja erinacea). Fish Shellfish Immunol, 25, 188-190.


[11]   Cotter MJ, Norman KE, Hellewell PG, Ridger VC (2001) A novel method for isolation of neutrophils from murine blood using negative immunomagnetic separation. Am J Pathol, 159, 473-481.

[12]   Burger DE, Wang L, Ban L, Okubo Y, Kühtreiber WM, et al. (2011) Novel automated blood separations validate whole cell biomarkers. PLoS One 6: e22430.

[13]   Ward DM (2011) Conventional apheresis therapies: a review. J Clin Apher, 26, 230-238.


[15]   Ghosh S, Kaplan G, Panaccione R (2009) Granulocyte-monocyte apheresis for the treatment of ulcerative colitis-is this the end of the road ? Nat Clin Pract Gastroenterol Hepatol, 6, 6-7.

[16]   Adacolumn

[17]   Danese S, Angelucci E, Marconi S, Repici A, Malesci A (2008) Oral beclomethasone: a review of its use in inflammatory bowel disease. Mini Rev Med Chem, 8, 1459-1463.


[19]   Dzik S (1993) Leukodepletion blood filters: filter design and mechanisms of leukocyte removal. Transfus Med Rev, 7, 65-77.

[20]   Ferrer F, Rivera J, Corral J, González-Conejero R, Vicente V (2000) Evaluation of leukocyte-depleted platelet concentrates obtained by in-line filtration. Vox Sang, 78, 235-241.

[21]   Mascaretti L, Baggi L, Riva M, Proserpio P, Dassi M, et al. (2002) Lymphocyte subsets in inline filtered packed red blood cell units: comparison between low and high spin procedures. Transfus Apher Sci, 26, 167-174.

[22]   Okuzaki D, Kimura S, Yabuta N, Ohmine T, Nojima H (2011) LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood. BMC Clin Pathol, 11, 9.