Skip to main content
SpringerLink
Log in

To disaggregate or not to disaggregate, injury and cell disaggregation, transient or permanent?

  • Published:
In Vitro Aims and scope Submit manuscript

Summary

Methods for disaggregating tissues to obtain cells for primary cultures are reviewed and evaluated. The procedures include dissociation with crude and purified enzymes (singly and in various combinations), depletion of cations, and manipulation of pH and osmolality.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Rous, P., and F. S. Jones. 1916. A method for obtaining suspensions of living cells from the fixed tissues, and for the plating out of individual cells. J. Exp. Med. 23: 549–555.

    Article  CAS  PubMed  Google Scholar 

  2. Vogelaar, J. P. M., and E. Erlichman. 1934. Some remarks on the growth of human fibroblasts in media containing copper. Am. J. Cancer 22: 66–75.

    CAS  Google Scholar 

  3. Medawar, P. B. 1941. Sheets of pure epidermal epithelium from human skin. Nature 148: 783.

    Google Scholar 

  4. Willmer, E. N. 1945. Growth and form in tissue culture. In: W. E. Le Gros Clark (Ed.),Essays on Growth and Form. Oxford University Press, New York, pp. 264–294.

    Google Scholar 

  5. Dulbecco, R. 1952. Production of plaques in monolayer tissue cultures by single particles of an animal virus. Proc. Natl. Acad. Sci. U.S.A. 38: 747–752.

    Article  PubMed  CAS  Google Scholar 

  6. Moscona, A. 1952. Cell suspensions from organ rudiments of chick embryos. Exp. Cell Res. 3: 535–539.

    Article  CAS  Google Scholar 

  7. Scherer, W. F., J. T. Syverton, and G. O. Gey. 1953. Studies on the propagation in vitro of poliomyelitis viruses. IV. Viral multiplication in a stable strain of human malignant epithelial cells (strain HeLa) derived from an epidermal carcinoma of the cervix. J. Exp. Med. 97: 695–710.

    Article  PubMed  CAS  Google Scholar 

  8. Zeidman, I. 1947. Chemical factors in the mutual adhesiveness of epithelial cells. Cancer Res. 7: 386–389.

    CAS  PubMed  Google Scholar 

  9. Rinaldini, L. M. J. 1958. The isolation of living cells from animal tissues. Int. Rev. Cytol. 7: 587–647.

    CAS  Google Scholar 

  10. Rinaldini, L. M. 1954. A quantitative method for cell culture. J. Physiol. (Lond.) 123: 20P-21P.

    Google Scholar 

  11. Rinaldini, L. M. 1954. A quantitative method for growing animal cellsin vitro. Nature 173: 1134–1135.

    Article  PubMed  CAS  Google Scholar 

  12. Norrby, K., F. Knutson, and P. M. Lundin. 1966. On the single cell state in enzymatically produced tumor cell suspensions. Exp. Cell Res. 44: 421–428.

    Article  PubMed  CAS  Google Scholar 

  13. Pine, L., G. C. Taylor, D. M. Miller, G. Bradley, and H. R. Wetmore. 1969. Comparison of good and bad lots of trypsin used in the production of primary monkey kidney cells. A definition of the problem and comparison of certain enzymatic characteristics. Cytobios 2: 197–207.

    Google Scholar 

  14. Kasten, F. 1973. Preparation of primary cultures. Chap. 10. Mammalian myocardial cells. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 72–81.

    Google Scholar 

  15. Glaeser, R. M., J. E. Richmond, and P. W. Todd. 1968. Histotypic self-organization by trypsin-dissociated and EDTA-dissociated chick embryo cells. Exp. Cell Res. 52: 71–85.

    Article  PubMed  CAS  Google Scholar 

  16. Shipman, C. 1973. Primary tissue dissociation. Chap. 2. Trypsin. A. Mammalian tissues In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 5–8.

    Google Scholar 

  17. Hull, R. N., G. Weaver, and W. R. Cherry. 1973. Preparation of primary tissues. Chap. 14. Monkey kidney cells. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 97–101.

    Google Scholar 

  18. Rappaport, C. 1956. Trypsinization of monkey-kidney tissue: an automatic, method for the preparation of cell suspensions. Bull. W.H.O. 14: 147–166.

    PubMed  CAS  Google Scholar 

  19. Parker, R. C. 1961.Methods of Tissue Culture. 3rd. ed. Hoeber, New York, pp. 121–130.

    Google Scholar 

  20. Paul, J. 1970.Cell and Tissue Culture, 4th ed. E. and S. Livingstone, Ltd., Edinburgh, pp. 203–212.

    Google Scholar 

  21. Kammer, H. 1969. Cell dispersal methods for increasing yield from animal tissues. Appl. Microbiol. 17: 524–527.

    PubMed  CAS  Google Scholar 

  22. Lasfargues, E. Y., and L. Ozzello. 1958. Cultivation of human breast carcinomas. J. Natl. Cancer Inst. 21: 1131–1147.

    PubMed  CAS  Google Scholar 

  23. Lasfargues, E. Y. 1973. Preparation of primary cultures. Chap. 3. Human mammary tumors. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 45–50.

    Google Scholar 

  24. Moscona, A., and H. Moscona. 1952. The dissociation and aggregation of cells from organ rudiments of the early chick embryo. J. Anat. 86: 287–301.

    PubMed  CAS  Google Scholar 

  25. Edwards, G. A., and J. Fogh. 1959. Micromorphologic changes in human amnion cells during trypsinization. Cancer Res. 19: 608–611.

    PubMed  CAS  Google Scholar 

  26. Allen, A., and C. Snow. 1970. The effect of trypsin or ethylenediamine tetraacetate on the surface of cells in tissue culture. Biochem. J. 117: 32P.

  27. Pumper, R. W., P. Fagan, and W. G. Taylor. 1971. Trypsin sensitivity of mammalian cells grown in a serum-free medium. In Vitro 6: 266–268.

    PubMed  CAS  Google Scholar 

  28. Cook, G. M. W., D. H. Heard, and G. V. F. Seaman. 1960. A sialomucopeptide liberated by trypsin from the human erythrocyte. Nature 138:1011–1012.

    Article  Google Scholar 

  29. Winzler, R. J. 1969. The association of a glycoprotein component of erythrocyte stroma with the cell membrane. In: R. T. Smith and R. A. Good (Eds.),Cellular Recognition. Appleton-Century-Crofts, New York, pp. 11–18.

    Google Scholar 

  30. Laws, J. O., and L. H. Strickland. 1961. The adhesion of liver cells. Exp. Cell. Res. 24: 240–254.

    Article  PubMed  CAS  Google Scholar 

  31. Weiser, R. S., E. Heise, K. McIvor, S. H. Han, and G. A. Granger. 1969.In vitro activities of immune macrophages. In: R. T. Smith and R. A. Good (Eds.),Cellular Recognition. Appleton-Century-Crofts, New York, pp. 215–220.

    Google Scholar 

  32. Snow, C., and A. Allen. 1970. The release of radioactive nucleic acids and mucoproteins by trypsin and ethylenediaminetetraacetate treatment of baby-hamster cells in tissue culture. Biochem. J. 119: 707–714.

    PubMed  CAS  Google Scholar 

  33. Hosick, H. L., and R. Strohman. 1971. Changes in ribosome-polyribosome balance in chick muscle cells during tissue dissociation, development in culture, and exposure to simplified culture medium. J. Cell. Physiol. 77: 145–156.

    Article  PubMed  CAS  Google Scholar 

  34. Hodges, G. M., D. C. Livingston, and L. M. Franks. 1973. The localization of trypsin in cultured mammalian cells. J. Cell Sci. 12: 887–902.

    PubMed  CAS  Google Scholar 

  35. Scheie, P., and H. Dalen. 1969. Surface features of cultured mammalian cells.Proceedings of the Second Annual Scanning Electron Microscope Symposium, 1969. Illinois Institute of Technology Press, Chicago.

    Google Scholar 

  36. Dalen, H., and P. O. Scheie. 1968. Two types of long microextensions from cultivated liver cells. Exp. Cell Res. 53: 670–673.

    Article  Google Scholar 

  37. Dalen, H., and P. Scheie. 1969. Microextensions on Chang's liver cells as observed throughout their division cycle. Exp. Cell Res. 57: 351–358.

    Article  PubMed  CAS  Google Scholar 

  38. Dalen, H., and P. W. Todd. 1971. Surface morphology of trypsinized human cells in vitro. Exp. Cell Res. 66: 353–361.

    Article  PubMed  CAS  Google Scholar 

  39. Sinclair, R., R. A. Reid, and P. Mitchell. 1963. Culture of strain L cells in suspension: replacement of polymer by traces of trypsin in a defined medium. Nature 197: 982–984.

    Article  PubMed  CAS  Google Scholar 

  40. Day, T. M., and A. H. Maddy. 1968. The turnover of the fibroblast surface. Ex. Cell Res. 53: 665–669.

    Article  CAS  Google Scholar 

  41. Hebb, C. R., and M.-Y. Wang Chu. 1960. Reversible injury of L-strain mouse cells by trypsin. Exp. Cell Res. 20: 453–457.

    Article  CAS  Google Scholar 

  42. Poste, G. 1971. Tissue dissociation with proteolytic enzymes. Exp. Cell Res. 65: 359–367.

    Article  PubMed  CAS  Google Scholar 

  43. Auerbach, R., and C. Grobstein. 1958. Inductive interaction of embryonic tissues after dissociation and reaggregation. Exp. Cell Res. 15: 384–397.

    Article  PubMed  CAS  Google Scholar 

  44. Green, N. M., and H. Neurath. 1953. The effects of divalent cations on trypsin. J. Biol. Chem. 204: 379–390.

    PubMed  CAS  Google Scholar 

  45. Pine, L., G. Bradley, and D. Miller. 1969. Collagenolytic activity and the comparison of good and bad lots of commercial trypsin in the preparation of primary monkey kidney cells. Cytobios 4: 347–367.

    Google Scholar 

  46. Oakley, C. L., G. H. Warrack, and W. E. Van Heyningen. 1946. The collagenase (κ toxin) ofCl. welchii type A. J. Pathol. Bacteriol. 58: 229–235.

    Article  CAS  Google Scholar 

  47. Bernfield, M. R., and N. K. Wessells. 1970. Intra- and extracellular control of epithelial morphogenesis. Dev. Biol. 4: 195–249.

    Google Scholar 

  48. Bernfield, M. R., S. D. Banerjee, and R. H. Cohn. 1972. Dependence of salivary epithelial morphology and branching morphogenesis upon acid mucopolysaccharide-protein (proteoglycan) at the epithelial surface. J. Cell Biol. 52: 674–689.

    Article  PubMed  CAS  Google Scholar 

  49. Kono, T. 1968. Purification and partial characterization of collagenolytic enzymes fromClostridium histolyticum. Biochemistry 7: 1106–1114.

    Article  PubMed  CAS  Google Scholar 

  50. Kono, T. 1969. Roles of collagenases and other proteolytic enzymes in the dispersal of animal tissue. Biochim. Biophys. Acta 178: 397–400.

    PubMed  CAS  Google Scholar 

  51. Bidwell, E., and W. E. Van Heyningen 1948. The biochemistry of the gas gangrene toxins. 5. The κ-toxin (collagenase) ofClostridium welchii. Biochem. J. 42: 140–151.

    PubMed  CAS  Google Scholar 

  52. Lasfargues, E. Y., and D. H. Moore. 1971. A method for the continuous cultivation of mammary epithelium. In Vitro 7: 21–25.

    PubMed  CAS  Google Scholar 

  53. Grant, N. H., and H. E. Alburn. 1960. Collagen solubilization by mammalian proteinases. Arch. Biochem. Biophys. 89: 262–270.

    Article  PubMed  CAS  Google Scholar 

  54. Houck, J. C., and Y. M. Patel. 1962. The collagenolytic activity of pancreas. Ann. N. Y. Acad. Sci. 93: 331–350.

    Article  CAS  Google Scholar 

  55. Hilfer, S. R. 1973. Primary tissue dissociation. Chap. 3. Collagenase treatment of chick heart and thyroid. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 16–20.

    Google Scholar 

  56. Grobstein, C., and J. Cohen. 1965. Collagenase: effect on the morphogenesis of embryonic salivary epithelium in vitro. Science 150: 626–628.

    Article  PubMed  CAS  Google Scholar 

  57. Wessells, N. K., and J. H. Cohen. 1968. Effects of collagenase on developing epithelialin vitro: lung, ureteric bud, and pancreas. Dev. Biol. 18: 294–309.

    Article  PubMed  CAS  Google Scholar 

  58. Zendzian, E. N., and E. A. Barnard. 1967. Distribution of pancreatic ribonuclease, chymotrypsin, and trypsin in vertebrates. Arch. Biochem. Biophys. 122: 699–713.

    Article  PubMed  CAS  Google Scholar 

  59. Naughton, M. A., and F. Sanger. 1960. Purification and specificity of pancreatic, elastase. Biochem. J. 78: 156–163.

    Google Scholar 

  60. Shotton, D. M. 1970. Elastase. Methods in Enzymol. 19: 113–140.

    Article  CAS  Google Scholar 

  61. Rinaldini, L. M. 1959. An improved method for the isolation and quantitative cultivation of embryonic cells. Exp. Cell Res. 16: 477–505.

    Article  PubMed  CAS  Google Scholar 

  62. Levinson, C., and J. W. Green. 1965. Cellular injury resulting from tissue disaggregation. Exp. Cell Res. 39: 309–317.

    Article  PubMed  CAS  Google Scholar 

  63. Phillips, H. J. 1972. Dissociation of single cells from lung or kidney with elastase. In Vitro 8: 101–105.

    PubMed  CAS  Google Scholar 

  64. Day, A. J., and G. K. Wilkinson. 1967. Incorporation of14C-labeled acetate into lipid by isolated foam cells and by atherosclerotic arterial intima. Circ. Res. 21: 593–600.

    PubMed  CAS  Google Scholar 

  65. Baló, J., and I. Banga. 1949. Elastase and elastase-inhibitor. Nature 164: 491.

    PubMed  Google Scholar 

  66. Gwatkin, R. B. L., 1963. Studies on the zona pellucida of the mouse egg. J. Reprod. Fertil. 6: 325.

    Google Scholar 

  67. Gwatkin, R. B. L. 1963. Effect of viruses on early mammalian development. I. Action of Mengo encephalitis virus on mouse ova cultivated in vitro. Proc. Natl. Acad. Sci. U.S.A.50: 576–581.

    Article  PubMed  CAS  Google Scholar 

  68. Gwatkin, R. B. L. 1964. Effects of enzymes and acidity on the zona pellucida of the mouse egg before and after fertilization. J. Reprod. Fertil7: 99–105.

    Article  PubMed  CAS  Google Scholar 

  69. Mintz, B. 1962. Experimental study of the developing mammalian egg: removal of the zona pellucida. Science 138: 594–595.

    Article  PubMed  CAS  Google Scholar 

  70. Edwards, R. G. 1964. Cleavage of one- and two-celled rabbit eggsin vitro after removal of the zona pellucida. J. Reprod. Fertil.7: 413–415.

    Article  PubMed  CAS  Google Scholar 

  71. Smithberg, M. 1953. The effect of different proteolytic enzymes on the zona pellucida of mouse ova. Anat. Rec.117: 554.

    Google Scholar 

  72. Gwatkin, R. B. L., and J. L. Thomson. 1964. A new method for dispersing the cells of mammalian tissues. Nature 201: 1242–1243.

    Article  PubMed  CAS  Google Scholar 

  73. Bowman, P., and A. McLaren. 1970. Viability and growth of mouse embryos afterin vitro culture and fusion. J. Embryol. Exp. Morphol. 23: 693–704.

    PubMed  CAS  Google Scholar 

  74. Bowman, P., and A. McLaren. 1970. The reaction of the mouse blastocyst and its zona pellucida to enzymesin vitro. J. Embryol. Exp. Morphol. 24: 331–334.

    PubMed  CAS  Google Scholar 

  75. Wilson, B. W., and T. L. Lau. 1963. Dissociation and cultivation of chick embryo cells with an Actinomycete protease. Proc. Soc. Exp. Biol. Med. 114: 649–651.

    PubMed  CAS  Google Scholar 

  76. Kahn, J., M. J. Ashwood-Smith, and D. M. Robinson. 1965. A cautionary note on the use of pronase in tissue culture. Exp. Cell Res. 40: 445–446.

    Article  PubMed  CAS  Google Scholar 

  77. Sullivan, J. C., and I. A. Schafer. 1966. Survival of pronase-treated cells in tissue culture. Exp. Cell Res. 43: 676.

    Article  PubMed  CAS  Google Scholar 

  78. Weinstein, D. 1966. Comparison of pronase and trypsin for detachment of human cells during serial cultivation. Exp. Cell Res. 43: 234–236.

    Article  PubMed  CAS  Google Scholar 

  79. Foley, J. F., and B. Aftonomos. 1970. The use of pronase in tissue culture: a comparison with trypsin. J. Cell. Physiol. 75: 159–162.

    Article  PubMed  CAS  Google Scholar 

  80. Houba, V. 1967. The use of pronase for dispersing cells. Experientia 23: 572.

    Article  PubMed  CAS  Google Scholar 

  81. Gwatkin, R. B. L. 1973. Primary tissue dissociation. Chap. 1. Pronase. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 3–5.

    Google Scholar 

  82. Kalant, H., and F. G. Young. 1957. Metabolic behaviour of isolated liver and kidney cells. Nature 179: 816–817.

    Article  PubMed  CAS  Google Scholar 

  83. Curtis, A. S. G. 1967.The Cell Surface: Its Molecular Role in Morphogenesis. Logos Press, London.

    Google Scholar 

  84. Armstrong, P. B. 1966. On the role of metal cations in cellular adhesion: effect on cell surface charge. J. Exp. Zool. 163: 99–109.

    Article  PubMed  CAS  Google Scholar 

  85. Anderson, N. G. 1953. The mass isolation of whole cells from rat liver. Science 117: 627–628.

    Article  PubMed  CAS  Google Scholar 

  86. Zwilling, E. 1954. Dissociation of chick embryo cells by means of chelating compound. Science 120: 219.

    Article  PubMed  CAS  Google Scholar 

  87. Coman, D. R. 1954. Cellular adhesiveness in relation to the invasiveness of cancer: electron microscopy of liver perfused with a chelating agent. Cancer Res. 14: 519–521.

    PubMed  CAS  Google Scholar 

  88. St. George, S., M. Friedman, and S. O. Byers. 1954. Mass separation of reticuloendothelial and parenchymal cells of rat's liver. Science 120: 463–464.

    Article  PubMed  CAS  Google Scholar 

  89. Melnick, J. L., C. Rappaport, D. Banker, and P. Bhatt. 1955. Stabilized suspensions of monkey kidney cells suitable for intercontinental shipping. Proc. Soc. Exp. Biol. Med. 88: 676–678.

    PubMed  CAS  Google Scholar 

  90. Laws, J. O., and L. H. Strickland. 1956. Metabolism of isolated liver cells. Nature 178: 309–310.

    Article  PubMed  CAS  Google Scholar 

  91. Moscona, A., O. A. Trowell, and E. N. Willmer. 1965. Methods. 2. Chemical dissociation. Chap. 2. In: E. N. Willmer (Ed.),Cells and Tissues in Culture. Vol. 1. Academic Press, New York, pp. 52–54.

    Google Scholar 

  92. Ontko, J. A. 1967. Chylomicron, free fatty acid and ketone body metabolism of isolated liver cells and liver homogenates. Biochim. Biophys. Acta 137: 13–22.

    PubMed  CAS  Google Scholar 

  93. Moscona, A. A., and M. H. Moscona. 1967. Comparison of aggregation of embryonic cells dissociated with trypsin or Versene. Exp. Cell Res. 45: 239–243.

    Article  PubMed  CAS  Google Scholar 

  94. Easty, G. C., and V. Mutolo. 1960. The nature of the intercellular material of adult mammalian tissues. Exp. Cell Res. 21: 374–385.

    Article  PubMed  CAS  Google Scholar 

  95. Montes de Oca, H. 1973. Primary tissue dissociation. Chap. 2. Trypsin. B. High yield method for kidney tissue. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 8–12.

    Google Scholar 

  96. Schmid, R. W., and C. N. Reilley. 1957. New complexon for titration of calcium in the presence of magnesium. Anal. Chem. 29: 264–268.

    Article  CAS  Google Scholar 

  97. Paterson, B., and R. C. Strohman. 1972. Myosin synthesis in cultures of differentiating chicken embryo skeletal muscle. Dev. Biol. 29: 113–138.

    Article  PubMed  CAS  Google Scholar 

  98. Robb, J. A. 1973. Single cell isolation and cloning. Chap. 5. Microtest plates. B. Replica plating. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 370–374.

    Google Scholar 

  99. Kleinschuster, S. J., and A. A. Moscona. 1972. Interactions of embryonic and fetal neural retina cells with carbohydrate-binding phytoagglutinins: cell surface changes with differentiation. Exp. Cell Res. 70: 397–410.

    Article  PubMed  CAS  Google Scholar 

  100. Wallick, E. T., J. C. Allen, and A. Schwartz. 1973. Differential effects of metal chelators on Na+, K+-ATPase activity. Arch. Biochem. Biophys. 158: 149–153.

    Article  PubMed  CAS  Google Scholar 

  101. Bygrave, F. L. 1967. The ionic environment and metabolic control. Nature 214: 667–671.

    Article  PubMed  CAS  Google Scholar 

  102. Harris, C. C., and C. A. Leone. 1966. Some effects of EDTA and tetraphenylboron on the ultra structure of mitochondria in mouse liver cells. J. Cell Biol. 28: 405–408.

    Article  PubMed  CAS  Google Scholar 

  103. Mazia, D. 1954. The particulate organization of the chromosome. Proc. Natl. Acad. Sci. U.S.A. 40: 521–527.

    Article  PubMed  CAS  Google Scholar 

  104. Rappaport, C., and G. B. Howze. 1966. Dissociation of adult mouse liver by sodium tetraphenylboron, a potassium complexing agent. Proc. Soc. Exp. Biol. Med. 121: 1010–1016.

    PubMed  CAS  Google Scholar 

  105. Rappaport, C., and G. B. Howze. 1966. Further studies on the dissociation of adult mouse tissue. Proc. Soc. Exp. Biol. Med. 121: 1016–1021.

    PubMed  CAS  Google Scholar 

  106. Rappaport, C. 1966. Effect of temperature on dissociation of adult mouse liver with sodium tetraphenylboron. Proc. Soc. Exp. Biol. Med. 121: 1022–1025.

    PubMed  CAS  Google Scholar 

  107. Rappaport, C. 1966. Role of “intercellular” matrix in aggregation of mammalian cells. Proc. Soc. Exp. Biol. Med. 121: 1025–1028.

    PubMed  CAS  Google Scholar 

  108. Casanello, D., and L. E. Gerschenson. 1970. Some morphological and biochemical characteristics of isolated rat liver cells dissociated with sodium tetraphenylboron and cultured in suspension. Exp. Cell Res. 59: 283–290.

    Article  PubMed  CAS  Google Scholar 

  109. Friedman, T., and C. J. Epstein. 1967. The incorporation of3H-leucine into protein by tetraphenylboron- and citrate-dispersed rat liver parenchymal cells. Biochim. Biophys. Acta 138: 622–624.

    Google Scholar 

  110. Brown, J. M., and S. R. Hilfer. 1968. Effects of the dissociating agents collagenase, pronase, and tetraphenylboron on embryonic chick thyroid and heart cells. J. Cell Biol. 39: 18a.

    Google Scholar 

  111. Murthy, L., and H. G. Petering. 1969. A comparison of some biochemical properties of dissociated liver cells and slices. Proc. Soc. Exp. Biol. Med. 132: 931–935.

    PubMed  CAS  Google Scholar 

  112. Kerkof, P. R., S. Smith, H. T. Gagné, D. R. Pitelka, and S. Abraham. 1969. Nonviability of sodium tetraphenylboron-dissociated cells. Exp. Cell Res. 58: 445–448.

    Article  PubMed  CAS  Google Scholar 

  113. Parry, E. W. 1971. Membrane-bounded intranuclear structures in hepatocytes after exposure to sodium tetraphenylboron. J. Pathol. 104: 210–212.

    Article  PubMed  CAS  Google Scholar 

  114. Mookerjee, S. 1953. Experimental dissociation of cells from chick embryos. Nature 171: 746.

    Article  Google Scholar 

  115. Longmuir, I. S., and W. ap Rees. 1956. Preparation of cell suspensions from rat liver. Nature 177: 997.

    Article  PubMed  CAS  Google Scholar 

  116. McLimans, W. F., D. T. Mount, S. Bogitch, E. J. Crouse, G. Harris, and G. E. Moore. 1966. A controlled environment system for study of mammalian cell physiology. Ann. N.Y. Acad. Sci. 139: 190–213.

    Article  PubMed  CAS  Google Scholar 

  117. McLimans, W. F. 1969. Physiology of the cultured mammalian cells. In: G. L. Tritsch (Ed.),Axenic Mammalian Cell Reactions. Chap. 10. Marcel Dekker, Inc., New York, pp. 307–367.

    Google Scholar 

  118. Hoskins, J. M., G. G. Meynell, and F. K. Sanders. 1956. A comparison of methods for estimating the viable count of a suspension of tumour cells. Exp. Cell Res. 11: 297–305.

    Article  PubMed  CAS  Google Scholar 

  119. Phillips, H. J. 1973. Evaluation of culture dynamics. Chap. 3. Dye exclusion tests for cell viability. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 406–408.

    Google Scholar 

  120. Yip, D. K, and N. Auersperg. 1972. The dye-exclusion test for cell viability: persistence of differential staining following fixation. In Vitro 7: 323–329.

    PubMed  CAS  Google Scholar 

  121. Steinberg, M. S. 1962. The role of temperature in the control of aggregation of dissociated embryonic cells. Exp. Cell Res. 28: 1–10.

    Article  PubMed  CAS  Google Scholar 

  122. Suzangar, M., and J. A. Dickson. 1970. Biochemical studies on cells isolated from adult rat liver. Exp. Cell Res. 63: 353–364.

    Article  PubMed  CAS  Google Scholar 

  123. Gallai-Hatchard, J. J., and G. M. Gray. 1971. A method for obtaining a suspension of intact parenchymal cells from adult rat liver. J. Cell Sci. 8: 73–86.

    PubMed  CAS  Google Scholar 

  124. Weigand, K., M. Müller, J. Urban, and G. Schreiber. 1971. Intact endoplasmic reticulum and albumin synthesis in rat liver cell suspensions. Exp. Cell Res. 67: 27–32.

    Article  CAS  Google Scholar 

  125. Laufs, R., and W. Walker. 1970. Metabolic and virologic studies in primate liver organ culture. Proc. Soc. Exp. Biol. Med. 133: 1006–1017.

    PubMed  CAS  Google Scholar 

  126. Frimmer, M., and W. Kreh. 1971. The role of oxidative and glycolytic mechanisms in cation transport of the isolated rat liver perfused with an erythrocyte-free medium. Naunyn Schmiederbergs. Arch. Pharmacol. 268: 119–121.

    CAS  Google Scholar 

  127. Plagemann, P. G. W., and H. E. Swim. 1966. Propagation of lactic dehydrogenase-elevating virus in cell culture. Proc. Soc. Exp. Biol. Med. 121: 1147–1152.

    PubMed  CAS  Google Scholar 

  128. Rutsky, L. P., W. G. Taylor, and R. W. Pumper. 1971. In vitro stimulation of rat liver cells by partial hepatectomy serum. In Vitro 7: 1–5.

    Google Scholar 

  129. Paradisi, F., and F. Piccinino. 1968. Propagation of mouse hepatitis virus (MHV-3) in monolayer cells cultures from liver of newborn mice. Experientia 24: 373–374.

    Article  PubMed  CAS  Google Scholar 

  130. Pisano, J. C., J. P. Filkins, and N. R. DiLuzio. 1968. Phagocytic and metabolic activities of isolated rat Kupffer cells. Proc. Soc. Exp. Biol. Med. 128: 917–922.

    PubMed  CAS  Google Scholar 

  131. Coon, H. G. 1968. Clonal cultures of differentiated rat liver cells. J. Cell Biol. 39: 29A.

    Google Scholar 

  132. Kaighn, M. E., and A. M. Prince. 1971. Production of albumin and other serum proteins by clonal cultures of normal human liver. Proc. Natl. Acad. Sci. U.S.A. 68: 2396–2400.

    Article  PubMed  CAS  Google Scholar 

  133. Howard, R. B., A. K. Christensen, F. A. Gibbs, and L. A. Pesch. 1967. The enzymatic preparation of isolated intact parenchymal cells from rat liver. J. Cell Biol. 35: 675–684.

    Article  PubMed  CAS  Google Scholar 

  134. Howard, R. B. 1968. Respiratory activity of intact, isolated parenchymal cells from rat liver. J. Biol. Chem. 243: 3105–3109.

    PubMed  CAS  Google Scholar 

  135. Berry, M. N., and D. S. Friend. 1969. High-yield preparation of isolated rat liver parenchymal cells. J. Cell Biol. 43: 506–520.

    Article  PubMed  CAS  Google Scholar 

  136. Hook, G. E. R., K. S. Dodgson, and F. A. Rose. 1970. Lysosomal arylsulfatases A and B of rat liver. Biochem. J. 119: 28P-29P.

    PubMed  CAS  Google Scholar 

  137. Cooper, B., and S. Margolis. 1971. Inhibition of lipid synthesis in isolated rat hepatocytes by serum lipoproteins. J. Lipid Res. 12: 731–739.

    PubMed  CAS  Google Scholar 

  138. Iype, P. T. 1971. Cultures from adult rat liver cells. I. Establishment of monolayer cell cultures from normal liver. J. Cell. Physiol. 78: 281–288.

    Article  PubMed  CAS  Google Scholar 

  139. Curtis, J. C., P. Lawner, and D. Minasian. 1968. Effect of the concentration of collagenase upon the dispersion of parenchymal cells of mouse liver. J. Cell Biol. 39: 30A.

    Google Scholar 

  140. Leffert, H. L., and D. Paul. 1973. Studies on primary cultures of differentiated fetal liver cells. J. Cell Biol. 52: 559–568.

    Article  Google Scholar 

  141. Fuzi, M., M. Toth, and J. Borsy. 1960. Isolation of cells with elastase during tissue culture. Zentralbl. Bakteriol [Orig. A] 177: 540–546. (Ref. from Chem. Abstr. 55: 1776.)

    Google Scholar 

  142. Hommes, F. A., M. I. Draisma, and I. Molenaar. 1970. Preparation and some properties of isolated rat liver cells. Biochim. Biophys. Acta 222: 361–371.

    PubMed  CAS  Google Scholar 

  143. Jacob, S. T., and P. M. Bhargava. 1962. A new method for the preparation of liver cell suspensions. Exp. Cell. Res. 27: 453–467.

    Article  PubMed  CAS  Google Scholar 

  144. Bhargava, P. M. 1970. Intercellular control of intracellular metabolic activity. In: G. E. W. Wolstenholme and J. Knight (Eds.),Control Processes in Multicellular Organisms. CIBA, symposium, J. and A. Churchill, Ltd., London, pp. 158–177.

    Google Scholar 

  145. Jacobs, M. H. 1928. The exchange of materials between the erythrocyte and its surroundings. Harvey Lect. 22: 146–164.

    Google Scholar 

  146. Wilson, T. H. 1954. Ionic permeability and osmotic swelling of cells. Science 120: 104–105.

    Article  PubMed  CAS  Google Scholar 

  147. Boyle, W. 1968. An extension of the51Cr-release assay for the estimation of mouse cytotoxins. Transplantation 6: 761–764.

    Article  PubMed  CAS  Google Scholar 

  148. Dickson, J. A. 1971. Metabolism of adult rat liver cells in filter-well culture. Exp. Cell Res. 64: 7–28.

    Article  Google Scholar 

  149. ap Gwynn, I., B. Jones, B. M. Jones, and R. B. Kemp. 1970. A modified perfusion technique for preparing isolated liver cells: an ultrastructural study. Cytobios 2: 181–191.

    CAS  Google Scholar 

  150. von Kraemer Bryn, M., and R. Oftebro. 1971. Pure primary cultures of adult rat liver macrophages. Exp. Cell Res. 69: 301–306.

    Article  Google Scholar 

  151. Lasfargues, E. Y. 1957. Cultivation and behavior in vitro of the normal mammary epithelium of the adult mouse. Anat. Rec. 127: 117–125.

    Article  PubMed  CAS  Google Scholar 

  152. Lasfargues, E. Y. 1957. Cultivation and behaviorin vitro of the normal mammary epithelium of the adult mouse. II. Observations on the secretory activity. Exp. Cell Res. 13: 553–562.

    Article  PubMed  CAS  Google Scholar 

  153. Wiepjes, G. J., and F. J. A. Prop. 1970. Improved method for preparation of single-cell suspensions from mammary glands of adult virgin mice. Exp. Cell Res. 61: 451–454.

    Article  PubMed  CAS  Google Scholar 

  154. Prop, F. J. A., and G. J. Wiepjes. 1973. Primary tissue dissociation. Chap. 4. Sequential enzyme treatment of mouse mammary gland. In: P. F. Kruse and M. K. Patterson (Eds.),Tissue Culture: Methods and Applications. Academic Press, New York, pp. 21–24.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Jackson Laboratory, 04609, Bar Harbor, Maine

    Charity Waymouth

Authors
  1. Charity Waymouth
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by National Institutes of Health Research Grant CA-11339 from the National Cancer Institute. The Jackson Laboratory is fully accredited by the American Association for Accreditation of Laboratory Animal Care. The experimental work on liver cell dissociation referred to was carried out by Alice T. Noyes, Patricia M. Ward, Ellen Akeson, and Susan Torrey.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Waymouth, C. To disaggregate or not to disaggregate, injury and cell disaggregation, transient or permanent?. In Vitro 10, 97–111 (1974). https://doi.org/10.1007/BF02615343

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02615343

Keywords

Search

Navigation