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Isolation of Human Blood Phagocytes by Counterflow Centrifugation Elutriation

  • Fabian J. Lionetti
  • Stephen M. Hunt
  • C. Robert Valeri
Part of the Biological Separations book series (BIOSEP)

Abstract

The principle of cell separation by counterflow centrifugation was first enunciated by Lindahl (1948) who derived an equation defining the position of particles in a centrifugal field opposed by fluid flowing in the centripetal direction. He designed a “counterstreaming” centrifuge which concentrated yeast particles in planes of equilibrium dependent upon the radius and density of the yeast and the viscosity and density of the medium (Lindahl, 1956). In an early study with the “counterstreaming” centrifuge eosinophilic leukocytes from horse blood were concentrated and the final cell suspension contained 20–30% (Lindahl and Lindahl, 1955). A significant advance was made by McEwen et al. (1968) who designed rotors for use with a standard preparative centrifuge. Their studies were largely responsible for the production of the simple equipment which makes counterflow cell separation generally available. Polystyrene micro spheres, yeasts, plant, and blood cells were resolved into subpopulations with diameters which ranged from 0–20 µm. In the first attempts to isolate leukocytes, whole blood with a ratio of 700 red cells to 1 leukocyte was concentrated to a ratio of 4:1. In a sample of whole blood in which 73% of the white cells consisted of granulocytes, counterflow centrifugation recovered 94% of them in the separation chambers. With a prototype rotor designed later, McEwen et al. (1971) achieved 99% recovery of leukocytes from malaria-infected monkey blood.

Keywords

Cell Separation Buffy Coat Fluorescein Diacetate Centrifugation Elutriation Separation Chamber 
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References

  1. Ackerman, S. K., and Douglas, S. D., 1978, Purification of human monocytes on microexudate-coated surfaces, J. Immunol 120: 1372.PubMedGoogle Scholar
  2. Bennett, W. E., and Cohn, Z. A. 1966, The isolation and selected properties of blood monocytes, J. Exp. Med. 123: 145.PubMedCrossRefGoogle Scholar
  3. Contreras, T. J., Hunt, S. M., Lionetti, F. J., and Valeri, C. R., 1978a, Preservation of human granulocytes. III. Liquid preservation studied by electronic sizing, Transfusion 18: 46.PubMedCrossRefGoogle Scholar
  4. Contreras, T. J., Jemionek, J. F., French, J. E., and Hartwig, V., 1978b, Liquid preservation of canine granulocytes obtained by counterflow centrifugation, Exp. Hematol. (Copenhagen) 6: 767.Google Scholar
  5. Contreras, T. J., Jemionek, J. F., French, J. E., and Hartwig, V., 1978b, Liquid preservation of canine granulocytes obtained by counterflow centrifugation, Exp. Hematol. (Copenhagen) 6: 767.Google Scholar
  6. Djerassi, I., Kim, J. S., and Suvansri, U., 1973, Harvesting human monocytes as a by-product of filtration-leukophoresis, Proc. Am. Assoc. Cancer Res. 14: 103 (Abstr.).Google Scholar
  7. Fogelman, A. M., Seager, J., Edwards, P. A. Kokom, M., and Popjak, G., 1977, Cholesterol biosynthesis in human lymphocytes, monocytes and granulocytes, Biochem. Biophys. Res. Commun. 76: 167.Google Scholar
  8. Grabske, R. J., 1978, Separating cell populations by elutriation. Fractions, Beckman Bull. 1: 1.Google Scholar
  9. Griffith, O. M., 1978, Separation of human T and B cells from human peripheral blood by centrifugal elutriation, Anal. Biochem. 87: 97.PubMedCrossRefGoogle Scholar
  10. Huestis, D. W., 1977, Production, storage and histocompatibility of granulocytes, in Blood Leukocytes Functions and Use in Therapy C. H. Hogman, K. Lindahl-Kiessling, and H. Higzell, eds.), Reklam and Katalogtryk, Uppsala, p. 55.Google Scholar
  11. Hunt, S. M., Lionett, F. J., and Valeri, C. R., 1980, Isolation and cryogenic preservation of human blood monocytes (in preparation).Google Scholar
  12. Jemionek, J. F., Contreras, T. J., Frenceh, J. E., and Hartwig, V., 1978a, Improved technique for increased granulocyte recovery from canine whole blood samples by counterflow centrifugation elutriation. I. In vitro analysis, Exp. Hematol. (Copenhagen) 6: 558.Google Scholar
  13. Jemionek, J. F., Contreras. T. J., French, J. F., and Shields, L. J., 1978b, Granulocyte isolation by counterflow centrifugation elutriation of canine blood obtained by continuous flow centrifugation leukopheresis, E.rp. Hematol. (Copenhagen) 6: 801.Google Scholar
  14. Jemionek, J. F., Contreras, T. C., French, J. E., and Shields, L. J., 1979, Technique for increased granulocyte recovery from human whole blood by counterflow centrifugation elutriation. I. In vitro analysis, Transfusion 19: 120.PubMedCrossRefGoogle Scholar
  15. Lindahl, P. E., 1948, Principle of a counterstreaming centrifuge for the separation of particles of different sizes, Nature 161: 648.PubMedCrossRefGoogle Scholar
  16. Lindahl, P. E., and Lindahl, K. M., 1955, On the concentration of eosinophile leukocytes, Experientia 11: 310.PubMedCrossRefGoogle Scholar
  17. Lindahl, P. E., 1956, On counterstreaming centrifugation in the separation of cells and cell fragments, Biochim. Biophys. Acta 21: 411.PubMedCrossRefGoogle Scholar
  18. Lionetti, F. J., Hunt, S. M., Lin, P. S., Kurtz, S. R., and Valeri, C. R., 1977, Preservation of human granulocytes. II. Characteristics of granulocytes obtained by counterflow centrifugation, Transfusion 17: 465.PubMedCrossRefGoogle Scholar
  19. Lionetti, F. J., Hunt, S. M., Mattaliano, R. J., and Valeri, C. R., 1978, In vitro studies of cryopreserved baboon granulocytes, Transfusion 18: 685.Google Scholar
  20. Loos, H., Blok-Schut, B., Van Doom, R., Hoksbergen, R., and Brutel de la Riviere, A., 1976a, A method for the recognition and separation of human blood monocytes on density gradients, Blood 48: 731.PubMedGoogle Scholar
  21. Loos, H., Blok-Schut, B., Kipp, B., Van Doom, R., and Meerhof, L., 19766, Size distribu- tion, electronic recognition and counting of human blood monocytes, Blood 48: 720.Google Scholar
  22. McEwen, C. R., Stallard, R. W. and Juhos, and E. Th.,1968, Separation of biological particles by centrifugal elutriations, Anal. Biochem. 23: 369.Google Scholar
  23. McEwen, C. R., Juhos, E. Th., Stallard, R. W., Schnell, J. V., Siddiqui, W. A., and Geiman, Q. M., 1971, Centrifugal elutriation for the removal of leukocytes from malaria-infected monkey blood, J. Parasitol. 57: 887.PubMedCrossRefGoogle Scholar
  24. Nunn, A. D., and Gagne, G., 1978, The recovery in small volume of cells from the Beckman JE-6 rotor, Transfusion 18: 599.PubMedCrossRefGoogle Scholar
  25. Persidsky, M. D., and Olson, L. S., 1978, Granulocyte separation by modified centrifugal elutriation system, Proc. Soc. Exp. Biol. Med. 157: 599.PubMedGoogle Scholar
  26. Pretlow, T. G., and Pretlow, T. P., 1979, Centrifugal elutriation (counterstreaming centrifugation) of cells, Cell Biophys. 1: 195.PubMedCrossRefGoogle Scholar
  27. Rabinowitz, Y., 1964, Separation of lymphocytes, polymorphonuclear leukocytes and monocytes on glass columns, including tissue culture observations, Blood 23: 811.PubMedGoogle Scholar
  28. Sanderson, R. J., and Bird, K. E., 1977, Cell separation by counterflow centrifugation, in Methods in Cell Biology ( D. M. Prescott, ed.), pp. 1–14, Academic Press, New York.Google Scholar
  29. Sanderson R. J., Bird, K. E., Palmer, N. F., and Brenman, J., 1976, Design principles for counterflow centrifugation cell separation chamber, Anal. Biochem. 71: 615.Google Scholar
  30. Sanderson, R. J., Sheppardson, F. T., Valter, A. E., and Talmadge, D. W., 1977, Isolation and enumeration of peripheral blood monocytes, J. Immunol. 118: 1409.PubMedGoogle Scholar
  31. Zeya, H. I. Keku, E., De Chatelet, L. R., Cooper, M. R., and Spun, C. L., 1978, Isolation of enzymatically homogeneous populations of human leukocytes, monocytes and granulocytes by zonal centrifugation, Am. J. Pathol. 90: 33.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • Fabian J. Lionetti
    • 1
  • Stephen M. Hunt
    • 1
  • C. Robert Valeri
    • 2
  1. 1.Center for Blood ResearchBostonUSA
  2. 2.Naval Blood Research LaboratoryBoston University Medical CenterBostonUSA

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