Separation of Tumor Cells from Fibroblasts

  • John A. Sykes

Abstract

It is well known that clinical and experimental microbiology made little progress until 1881, when Robert Koch described his method of preparing cultures on solid media. The use of solid media permitted rapid separation of different types of bacteria, and thus allowed the selected organisms to be grown out into pure cultures for further study. The art of tissue culture is somewhat akin to microbiology before the year 1881. It is very difficult to obtain “pure” cultures of selected cell types from most organs and tissues of man and animals. In studies of human tumors, this problem is particularly exasperating; the desired cell is the tumor cell, but the cell usually obtained is the fibroblast from the connective tissue elements associated with the tumor. The technique most widely used to obtain cultures of cells from organs and tissues of mammals utilizes controlled enzyme activity to break down the tissue fragment into a suspension of single cells or small groups of cells. The technique has proved effective for many tissues, particularly those derived from embryos or from organs without much fibrous stroma; however, adult tissues and those with abundant fibrous stroma provide a challenge and a stumbling block for the tissue culturist, who as it were wants the wheat, not the chaff and weeds. The methodology to be described, when tailored to the system being studied, has usually proved effective in separating a high proportion of tumor cells from the “contaminating” fibroblasts.

Keywords

Minimal Essential Medium Uninfected Cell Interface Zone Buoyant Density Ficoll Gradient 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Abeloff, M. D., Mangi, R. J., Pretlow, T. G., and Mardiney, M. R., 1970, Isolation of leukemic blasts from peripheral blood by density gradient centrifugation, J. Lab. Clin. Med. 75: 703–710.PubMedGoogle Scholar
  2. Ali, S. N., and Fletcher, K. A., 1971, Zonal centrifugation as a technique for the separation of malania-infected erythrocytes, Trans. R. Soc. Trop. Med. Hyg. 65: 4–5.PubMedCrossRefGoogle Scholar
  3. Boone, C. W., Harell, G. S., and Bone, H. E., 1968, The resolution of mixtures of viable mammalian cells into homogeneous fractions by zonal centrifugation, J. Cell Biol. 36: 369–378.PubMedCrossRefGoogle Scholar
  4. Böyum, A., 1968, Separation of leucocytes from blood and bone marrow, Scand. J. Clin. Lab. Invest. 21: 9–29.Google Scholar
  5. Britten, R. J., and Roberts, R. B., 1960, High resolution density gradient sedimentation analysis, Science 131: 32–33.PubMedCrossRefGoogle Scholar
  6. Gey, G. O., Coffman, W. D., and Kubicek, M. T., 1952, Tissue culture studies of the proliferative capacity of cervical carcinoma and normal epithelium, Cancer Res. 12: 264.Google Scholar
  7. Giorgi, P. P., 1971, Preparation of neurons and glial cells from rat brain by zonal centrifugation, Exp. Cell Res. 68: 273–282.PubMedCrossRefGoogle Scholar
  8. Holier, M., and Moller, K. M., 1958, A substance for aqueous density gradients, Exp. Cell Res. 15: 631–632.CrossRefGoogle Scholar
  9. Johnson, A. R., and Moran, M. C., 1966, Comparison of several methods for isolation of rat peritoneal mast cells, Proc. Soc. Exp. Biol. Med. 123: 886–889.PubMedCrossRefGoogle Scholar
  10. Johnson, L., Morrow, J., Kasten, F. H., and Dizerega, G., 1970, Isolation of viable neurons from embryonic spinal ganglia by centrifugation through albumin gradients, Exp. Cell Res. 63: 189–192.PubMedCrossRefGoogle Scholar
  11. Lasfargues, E. Y., and Ozzello, L., 1958, Cultivation of human breast carcinomas, J. Natl. Cancer Inst. 21: 1131–1 147.Google Scholar
  12. Leemann, U., Weiss, S., and Schmutz, E., 1971, A centrifugation technique for cytochemical preeparations, J. Histochem. Cytochem. 19: 758–760.PubMedCrossRefGoogle Scholar
  13. Mateyko, G. M., and Kopac, M. J., 1963, Cytophysical studies on living normal and neoplastic cells, Ann. N.Y. Acad. Sci. 105: 183–286.Google Scholar
  14. Pretlow, T. G., II, and Boone, C. W., 1968, Centrifugation of mammalian cells on gradients: A new rotor, Science 161: 911–913.PubMedCrossRefGoogle Scholar
  15. Rounds, D. E., 1970, A growth-modifying factor from cell lines of human malignant origin, Cancer Res. 30: 2847–2851.PubMedGoogle Scholar
  16. Schindler, R., Ramseier, L., Schaer, J. C., and Grieder, A., 1970, Studies on the division cycle of mammalian cells. 3. Preparation of synchronously dividing cells by isotonic sucrose gradient centrifugation, Exp. Cell Res. 59: 90–96.PubMedCrossRefGoogle Scholar
  17. Sykes, J. A., Grey, C. E., Scanlon, M., Young, L., and Dmochowski, L., 1964, Density gradient centrifugation studies of the Bittner virus, Texas Rep. Biol. Med. 22: 609–627.Google Scholar
  18. Sykes, J. A., Whitescarver, J., Jernstrom, P., Nolan, J. F., and Byatt, P., 1970a, Some properties of a new epithelial cell line of human origin, J. Natl. Cancer Inst. 45: 107–122.PubMedGoogle Scholar
  19. Sykes, J. A., Whitescarver, J., Briggs, L., and Anson, J. H., 1970b, Separation of tumor cells from fibroblasts with use of discontinuous density gradients, J. Natl. Cancer Inst. 44: 855–864.PubMedGoogle Scholar
  20. Uvnäs, B., and Thon, I. L., 1959, Isolation of “biologically intact” mask cells, Exp. Cell Res. 18: 512–520.PubMedCrossRefGoogle Scholar
  21. Walder, A. I., and Lumseth, J. B., 1963, A technic for separation of the cells of the gastric mucosa, Proc. Soc. Exp. Biol. Med. 112: 494–496.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1975

Authors and Affiliations

  • John A. Sykes
    • 1
  1. 1.Research Department, Southern California Cancer CenterCalifornia Hospital Medical CenterLos AngelesUSA

Personalised recommendations