Partial purification of human colonic carcinoma cells by sedimentation

Michael G Brattain, P. M. Kimball, T. G. Pretlow, A. M. Pitts

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

We have purified epithelial cells from human colonic tumours by velocity sedimentation in an isokinetic density gradient of Ficoll in tissue culture medium. In frozen sections of colonic carcinoma, histochemically demonstrable N-acetyl-beta-D-glucosaminidase (HDAG) was observed primarily in epithelial cells. We used this enzyme as a histochemical marker of epithelial cells. Initial suspensions of cells from colonic tumours suspended with 0-25% trypsin contained an average of 24% of the nucleated cells with HDAG. In the purest fraction obtained from gradient centrifugations, an average of 74% of the nucleated cells contained HDAG. After centrifugation, the quarter of the density gradient which contained the most rapidly sedimenting cells was purified 2-4-fold over that in the initial suspension. Cells in this zone of the gradient also gave rise to colonies in soft agar. Cells from initial suspension resulted in 15-25% as many colonies of 7 or more cells in cultures inoculated with the same number of nucleated cells. For the most part, cells obtained from the other zones of the gradient did not give rise to colonies in soft agar.

Original languageEnglish (US)
Pages (from-to)850-857
Number of pages8
JournalBritish journal of cancer
Volume35
Issue number6
DOIs
StatePublished - Jan 1 1977

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Carcinoma
Acetylglucosaminidase
Suspensions
Epithelial Cells
Centrifugation
Agar
Ficoll
Frozen Sections
Trypsin
Culture Media
Neoplasms
Cell Culture Techniques
Cell Count
Enzymes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Brattain, M. G., Kimball, P. M., Pretlow, T. G., & Pitts, A. M. (1977). Partial purification of human colonic carcinoma cells by sedimentation. British journal of cancer, 35(6), 850-857. https://doi.org/10.1038/bjc.1977.128

Partial purification of human colonic carcinoma cells by sedimentation. / Brattain, Michael G; Kimball, P. M.; Pretlow, T. G.; Pitts, A. M.

In: British journal of cancer, Vol. 35, No. 6, 01.01.1977, p. 850-857.

Research output: Contribution to journalArticle

Brattain, MG, Kimball, PM, Pretlow, TG & Pitts, AM 1977, 'Partial purification of human colonic carcinoma cells by sedimentation', British journal of cancer, vol. 35, no. 6, pp. 850-857. https://doi.org/10.1038/bjc.1977.128
Brattain, Michael G ; Kimball, P. M. ; Pretlow, T. G. ; Pitts, A. M. / Partial purification of human colonic carcinoma cells by sedimentation. In: British journal of cancer. 1977 ; Vol. 35, No. 6. pp. 850-857.
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