History and Overview of the In Vivo Diffusion Chamber (DC) Culture System

  • A. L. Carsten
Conference paper


The development of the modern DC system has been traced from the nearly 100-year-old concept for in vivo growth of isolated cells in a semipermeable container to the modern systems consisting of single- and multichambers with differing wall materials. Various applications of the system and methods of analysis are discussed. The many advantages and disadvantages of this system are considered as compared with other available culture systems.


Autologous Bone Marrow Host Animal Mouse Bone Marrow Plasma Clot Relative Biologic Effectiveness 
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  1. 1.
    Algire, G.H., Weaver, J.M., Prehn, R.T.: Growth of cells in vivo in diffusion chambers. I. Survival of homografts in immunized mice. J. Natl. Cancer Inst. 15, 493–501 (1954)PubMedGoogle Scholar
  2. 2.
    Ames, B.N., Durston, W.E., Yamasaki, E., Lee, F.D.: Carcinogens are mutagens: a simple test system combining liver homogenates for activation and bacteria for detection. Proc. Natl. Acad. Sci. USA 70, 2281–2285 (1973)PubMedCrossRefGoogle Scholar
  3. 3.
    Amos, D.: Transplantation of cells and tissues in diffusion chambers. In: Transplantation of tissues and cells. Billingham, R.E., Silvers, W.K. (eds.), pp. 69–80. Philadelphia: Wistar Inst. Press 1961Google Scholar
  4. 4.
    Amos, D.B., Wakefield, J.D.: Growth of mouse ascites tumor cells in diffusion chambers. I. Studies of growth rate of cells and of the rate of entry of antibody. J. Natl. Cancer Inst. 27, 658–670 (1958)Google Scholar
  5. 5.
    Benestad, H.B.: Formation of granulocytes and macrophages in diffusion chamber cultures of mouse blood leucocytes. Scand. J. Haematol. 7, 279–288 (1970)PubMedCrossRefGoogle Scholar
  6. 6.
    Benestad, H.B.: Blood-borne haemopoietic progenitor cells the mechanism of release and various parameters examined with the diffusion chamber technique. Norwegian Defence Research Establishment Report No. 61, Kjeller, Norway 1972Google Scholar
  7. 7.
    Benestad, H.B.: Cell kinetics in diffusion chambers: survival, resumption of proliferation, and maturation rate of murine haemopoietic cells. Cell Tissue Kinet. 5, 421–431 (1972)PubMedGoogle Scholar
  8. 8.
    Benestad, H.B., Breivik, H.: Properties of hematopoietic progenitor cells. Acta Physiol. Scand. 83, 389–398 (1971)PubMedCrossRefGoogle Scholar
  9. 9.
    Benestad, H.B., Iversen, J.G., Rolstad, B.: Immunoblast formation by recirculating and non-recirculating rat lymphocytes cultured in diffusion chambers. Scand. J. Haematol. 8/1, 32–43 (1971)Google Scholar
  10. 10.
    Ben-Ishay, Z.: Ultrastructural analysis of erythroid colonies in diffusion chambers in irradiated rats. Effects of cytosine arabinoside administration on donor rats. Scand. J. Haematol. 14, 369–375 (1975)PubMedCrossRefGoogle Scholar
  11. 11.
    Ben-Ishay, Z.: Selective eradication of rat bone marrow erythroid cells by administration of cytosine arabinoside. Observations on resumption phase of erythropoiesis. Scand. J. Haematol. 14, 361–368 (1975)PubMedCrossRefGoogle Scholar
  12. 12.
    Berman, I., Kaplan, H.S.: The cultivation of mouse bone marrow in vivo. Blood 14, 1040–1046 (1959)PubMedGoogle Scholar
  13. 13.
    Berman, I., Kaplan, H.S.: The functional capacity of mouse bone marrow cells growing in diffusion chambers. Exp. Cell. Res. 20, 238–239 (1960)PubMedCrossRefGoogle Scholar
  14. 14.
    Bisceglie, V.: Über die antineoplastische Immunität. II. Mitteilung. Über die Wachstumsfähigkeit der heterologen Geschwülste in erwachsenen Tieren nach Einpflanzung in Kollodium-Säckchen. Z. Krebsforsch. 40, 141–158 (1934)Google Scholar
  15. 15.
    Boecker, W.R., Bøyum, A., Carsten, A.L., Cronkite, E.P.: Human bone marrow (HBM) and blood (HPB) stem cell kinetics. Blood 38, 819 (1971)Google Scholar
  16. 16.
    Bøyum, A., Borgstrom, R.: The concentration of granulocytic stem cells in mouse bone marrow, determined with diffusion chamber technique. Scand. J. Haematol. 7, 294–303 (1970)PubMedCrossRefGoogle Scholar
  17. 17.
    Bøyum, A., Boecker, W., Carsten, A.L., Cronkite, E.P.: Proliferation of human bone marrow cells in diffusion chambers implanted into normal or irradiated mice. Blood 40, 163–173 (1972)PubMedGoogle Scholar
  18. 18.
    Bøyum, A., Carsten, A.L., Laerum, O.D., Cronkite, E.P.: Kinetics of cell proliferation of murine bone marrow cells cultured in diffusion chambers. Blood 40, 174–188 (1972)PubMedGoogle Scholar
  19. 19.
    Bøyum, A., Petersen, E.O., Laane, B., Carsten, A.L.: The diffusion chamber technique as progenitor (stem) cell assay. In: Hemopoiesis in culture. Robinson, W.A. (ed.), pp. 149–159. DHEW Publication No. (NIH) 74–205. Washington, D.C.: U.S. Gov. Print. Off. 1974Google Scholar
  20. 20.
    Bøyum, A., Carsten, A.L., Laerum, O.D.: Haematopoiesis measured by spleen colony and diffusion chamber techniques in mice treated with one or two injections of cyclophosphamide. Br. J. Haematol. 26, 605 (1974)PubMedCrossRefGoogle Scholar
  21. 21.
    Bøyum, A., Carsten, A.L., Chikkappa, G., Cook, L., Bullis, J., Honikel, L., Cronkite, E.P.: The r.b.e. of different-energy neutrons as determined by human bone-marrow cell-culture techniques. Int. J. Radiat. Biol. 34, 201–221 (1978)CrossRefGoogle Scholar
  22. 22.
    Breivik, H.: Haematopoietic stem cell content of murine bone marrow, spleen, and blood. Limiting dilution analysis of diffusion chamber cultures. J. Cell. Physiol. 78, 73–78 (1971)PubMedCrossRefGoogle Scholar
  23. 23.
    Breivik, H.: Response of multipotent (CFU) and granulocyte (diffusion chamber assay) progenitor cells and differentiating cells of murine haematopoietic tissues to a perturbation of the steady state. J. Cell. Physiol. 79 (2) 171–180 (1972)PubMedCrossRefGoogle Scholar
  24. 24.
    Breivik, H., Benestad, H.B.: Regulation of granulocyte and macrophage formation in diffusion chamber cultures of mouse haematopoietic cells. Exp. Cell Res. 70, 340–348 (1972)PubMedCrossRefGoogle Scholar
  25. 25.
    Breivik, H., Benestad, H.B., Bøyum, A.: Diffusion chamber and spleen colony assay of murine haematopoietic stem cells. J. Cell. Physiol. 78, 65–72 (1971)PubMedCrossRefGoogle Scholar
  26. 26.
    Brewen, M.G., Nettesheim, P., Jones, K.P.: A host-mediated assay for cytogenetic mutagenesis: preliminary data on the effect of methyl methanesulfonate. Mutat. Res. 10, 645–649 (1970)PubMedCrossRefGoogle Scholar
  27. 27.
    Capalbo, E.E., Makinodan, T.: Doubling time of mouse spleen cells during the latent and log phases of primary antibody response. J. Immunol. 92, 235–242 (1964)Google Scholar
  28. 28.
    Capalbo, E.E., Albright, J.F., Bennett, W.E.: Evaluation of the diffusion chamber culture technique for study of the morphological and functional characteristics of lymhoid cells during antibody production. J. Immunol. 92, 243–251 (1964)PubMedGoogle Scholar
  29. 29.
    Carsten, A.L., Bøyum, A., Boecker, W.: The culture of human and mouse hematopoietic stem cells in Millipore diffusion chambers. In: The Proceedings of a Workshop/Symposium on in In Vitro Culture of Hematopoietic Cells, Rijswijk, September 1971, Holland by Radiobiological Institute TNO, pp. 279–288Google Scholar
  30. 30.
    Carsten, A.L., Chanana, A.D., Chikkappa, G., Cronkite, E.P., Ohl, S.: An improved diffusion chamber (DC) for culture of hematopoietic cells. Proc. Soc. Exp. Biol. Med. 150, 107–109 (1975)PubMedGoogle Scholar
  31. 31.
    Carsten, A.L., Bond, V.P., Thompson, K.: The RBE of different energy neutrons as measured by the hematopoietic spleen-colony technique. Int. J. Radiat. Biol. 29, 65–70 (1976)CrossRefGoogle Scholar
  32. 32.
    Chikkappa, G., Boecker, W.R., Borner, G., Carsten, A.L., Conkling, K., Cook, L., Cronkite, E.P., Dunwoody, S.: Return of alkaline phosphatase in chronic myelocytic leukemia cells in diffusion chamber cultures. Proc. Soc. Exp. Biol. Med. 143, 212–217 (1973)PubMedGoogle Scholar
  33. 33.
    Chikkappa, G., Carsten, A.L., Chanana, A.D., Chandra, P., Cronkite, E.P.: Increased granulocytic, erythrocytic and megakaryocyte progenitors in myelofibrosis with myeloid metaplasia. Am. J. Haematol. 4, 121)131 (1978)CrossRefGoogle Scholar
  34. 34.
    Chikkappa, G., Carsten, A.L., Chanana, A.D., Cronkite, E.P.: Culture of normal human blood cells in diffusion chamber systems. I. Granulocytic cell survival and proliferation. Exp. Haematol. 6, 28–36 (1978)Google Scholar
  35. 35.
    Cronkite, E.P., Boecker, W., Carsten, A.L., Chikkappa, G., Joel, D., Laissue, J., Ohl, S.: The use of diffusion chamber cultures in the study of normal and leukemic cell proliferation in man. In: Hemopoiesis in culture. Robinsion, W.A. (ed.) pp. 185–199. DHEW Publication No. (NIH) 74–205. Washington, D.C.: U.S. Gov. Print. Off. 1974Google Scholar
  36. 36.
    Cronkite, E.P., Carsten, A.L., Chikkappa, G., Laissue, J.A., Ohl, S.: Culture of normal and leukemic cells in diffusion chambers. In: Advances in the biosciences 14: Workshop on Prognostic Factors in Human Acute Leukemia, Schloss Reisensberg, Germany, October 1-3, 1974. Fliedner, T.M., Perry, S. (eds.), pp. 273–286. New York: Pergamon Press 1975Google Scholar
  37. 37.
    Cronkite, E.P., Carsten, A.L., Cohen, R., Miller, M.E., Moccia, G.: The effects of humoral factors on amplification of nonrecognizable erythrocytic and granulocytic precursors. Blood Cells 5, 331–350 (1979)PubMedGoogle Scholar
  38. 38.
    Evgenjeva, T.P.: Heterotransplantation of human cancers to animals by means of diffusion chambers. Eur. J. Cancer 6, 533–535 (1970)PubMedCrossRefGoogle Scholar
  39. 39.
    Gerard, E., Carsten, A.L., Cronkite, E.P.: The proliferative potential of plasma clot erythroid colony forming cells in diffusion chambers. Blood Cells 4, 105–128 (1978)PubMedGoogle Scholar
  40. 40.
    Gordon, M.Y.: Quantitation of haemopoietic cells from normal and leukaemic RFM mice using an in vivo colony assay. Br. J. Cancer 30, 421–428 (1974)PubMedCrossRefGoogle Scholar
  41. 41.
    Gordon, M.Y., Blackett, N.M., Douglas, I.D.C.: Colony formation by human haemopoietic precursor cells cultured in semi-solid agar in diffusion chambers. Br. J. Haematol. 31, 103–110 (1975)PubMedCrossRefGoogle Scholar
  42. 42.
    Huang, C.C.: A modified host-mediated assay using cultured human lymphoid cells in diffusion chambers in mice and cytogenetic effects of cyclophosphamide. Environ. Res. 13, 267–277 (1977)PubMedCrossRefGoogle Scholar
  43. 43.
    Huang, C.C., Furukawa, M.: Sister chromatid exchanges in human lymphoid cell lines cultured in diffusion chambers in mice. A host-mediated assay system for mutagenicity. Exp. Cell Res. 111, 458–461 (1978)PubMedCrossRefGoogle Scholar
  44. 44.
    Jacobsen, N.: Chamber centrifugation: a harvesting technique for estimation of the growth of human haemopoietic cells in diffusion chambers. Br. J. Haematol. 29, 171–177 (1975)PubMedCrossRefGoogle Scholar
  45. 45.
    Johnson, L.I., Chan, P.C., LoBue, J., Monette, F.C., Gordon, A.S.: Cell cycle analysis of rat lymphocytes cultured with phytohemagglutinin in diffusion chambers. Exp. Cell Res. 47, 201–208 (1967)PubMedCrossRefGoogle Scholar
  46. 46.
    Johnson, L.I., LoBue, J., Chan, P.C., Monette, F.C., Rubin, A.D., Gordon, A.S., Dameshek, W.: Autoradiographic studies of human lymphocytes cultured in vivo. Proc. Soc. Exp. Biol. Med. 130, 675–679 (1969)PubMedGoogle Scholar
  47. 47.
    Laissue, J.A., Chanana, A.D., Cronkite, E.P., Joel, D.D., Pavelec, M.: Culture of autologous bone marrow in diffusion chambers: Effect of host irradiation. Blood 45, 417–425 (1975)PubMedGoogle Scholar
  48. 48.
    McVittie, T., McCarthy, K.: Increased proliferation of hematopoietic stem cells within diffusion chambers implanted into irradiated mice pretreated with cyclophosphamide. Radiat. Res. 59, 291 (1974)Google Scholar
  49. 49.
    Makinodan, T., Hoppe, I., Sado, T., Capalbo, E., Leonard, M.R.: The suppressive effect of supraoptimum doses of antigen on the secondary antibody-forming response of spleen cells cultured in cell-impermeable diffusion chambers. J. Immunol. 95, 466–479 (1965)PubMedGoogle Scholar
  50. 50.
    Meek, R.A., Carsten, A.L., Kelsch, J.J.: Growth of He La cells in diffusion chamber cultures in vivo. Cancer Res. 36, 2317–2320 (1976)Google Scholar
  51. 51.
    Metchnikoff, E.: Sur la lutte des cellules de l’organisme contre l’invasion des microbes. Ann. Inst. Pasteur 1, 321–336 (1887)Google Scholar
  52. 52.
    Miller, A.M., McGarry, M.P.: A diffusible stimulator of eosinophilopoiesis in vivo. Nature 262, 586 (1976)PubMedCrossRefGoogle Scholar
  53. 53.
    Miller, A.M., Colley, D.G., McGarry, M.P.: Spleen cells from Schistosoma mansoni infected mice produce diffusible stimulator eosinophilopoiesis in vivo. Nature 262, 586 (1976)PubMedCrossRefGoogle Scholar
  54. 54.
    Nettesheim, P., Makinodan, T.: Differentiation of lymphocytes undergoing an immune response in diffusion chambers. J. Immunol. 94, 868–876 (1965)PubMedGoogle Scholar
  55. 55.
    Nettesheim, P., Makinodan, T., Chadwick, C.J.: Improved diffusion chambers cultures for cytokinetic analysis of antibody response. Immunology 11, 427–439 (1966)PubMedGoogle Scholar
  56. 56.
    Niskanen, E., Tyler, W.S., Symann, M., Stohlman, F. Jr., Howard, D.: The effect of neutropenia on the cell cycle of granulocyte precursors in an in vivo culture system. Blood 43, 23–31 (1974)PubMedGoogle Scholar
  57. 57.
    Nocard, L., Roux, F.: Le microbe de la peripneumonie. Ann. Inst. Pasteur 12, 240–262 (1898)Google Scholar
  58. 58.
    Ohl, S., Carsten, A.L., Chanana, A.D., Chikkappa, G., Cronkite, E.P.: Increased erythrocytic and neutrophilic progenitors in myelofibrosis with myeloid metaplasia. Eur. J. Cancer 12, 131–135 (1976)PubMedCrossRefGoogle Scholar
  59. 59.
    Petrakis, N.L., Davis, M., Lucia, S.P.: The in vivo differentiation of human leukocytes into histiocytes, fibroblasts and fat cells in subcutaneous diffusion chambers. Blood 17, 109–118 (1961)PubMedGoogle Scholar
  60. 60.
    Quesenberry, P., Rappeport, J., Fountebuoni, A., Sullivan, R., Zuckerman, K., Ryan, M.: Inhibition of murine hematopoiesis by leukemia cells. N. Engl. J. Med. 299, 71–75 (1978)PubMedCrossRefGoogle Scholar
  61. 61.
    Rezzesi, F.D.: Eine Methode zur Züchtung der Gewebe in vivo. Arch. Exp. Zellforsch. 13, 258–281 (1932)Google Scholar
  62. 62.
    Shadduck, R.K., Carsten, A.L., Chikkappa, G., Cronkite, E.P., Gerard, E.: Inhibition of diffusion chamber (DC) granulopoiesis by anti-CSF serum. Proc. Soc. Exp. Biol. Med. 158, 542–549 (1978)PubMedGoogle Scholar
  63. 63.
    Shulman, L.H., Robinson, S.H.: Maintenance of hemopoietic stem cells in diffusion chamber culture. J. Lab. Clin. Med. 90, 581–588 (1977)PubMedGoogle Scholar
  64. 64.
    Steinberg, H.N., Handler, E.S., Handler, E.E.: Assessment of erythrocytic and granulocytic colony formation in an in vivo plasma clot diffusion chamber culture system. Blood 47, 1041–1051 (1976)PubMedGoogle Scholar
  65. 65.
    Symann, M.: Fetal hemopoiesis in diffusion chamber cultures. Thesis, Chatolic University of Louvain 1979Google Scholar
  66. 66.
    Tyler, W.S., Stohlman, F. Jr., Keane, J., Chovaniec, M., Howard, D.: Survival of murine hemopoietic cells in an in vivo culture system. Proc. Soc. Exp. Biol. Med. 146, 316–320 (1974)Google Scholar

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© Springer-Verlag Berlin Heidelberg 1980

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  • A. L. Carsten

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