Abstract
EMT-6/UW tumours were treated in vivo with X-rays, cyclophosphamide, or bleomycin. Cell survival was assayed in vitro following tumour disaggregation with trypsin or an enzyme cocktail (EC) consisting of pronase, collagenase and DNase which gives a 10-20 x higher cell yield. Surviving fraction was lower after cyclophosphamide treatment for cells isolated with EC than for cells prepared with trypsin. The opposite result was obtained with bleomycin; trypsin-isolated cells appeared more sensitive. In attempting to determine the basis for this discrepancy, it was found that both dissociation methods isolate a non-representative cell sample with fewer cells in DNA synthesis (12-13%) than in the original tumour (approximately 22%). The specific nature of the interaction between the injury caused by drug and enzyme remains to be elucidated.
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Selected References
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- Hodges G. M., Livingston D. C., Franks L. M. The localization of trypsin in cultured mammalian cells. J Cell Sci. 1973 May;12(3):887–902. doi: 10.1242/jcs.12.3.887. [DOI] [PubMed] [Google Scholar]
- Poste G. Tissue dissociation with proteolytic enzymes. Adsorption and activity of enzymes at the cell surface. Exp Cell Res. 1971 Apr;65(2):359–367. doi: 10.1016/0014-4827(71)90014-0. [DOI] [PubMed] [Google Scholar]
- RAUT HEBB C., WANG CHU M. Y. Reversible injury of L-strain mouse cells by trypsin. Exp Cell Res. 1960 Aug;20:453–457. doi: 10.1016/0014-4827(60)90174-9. [DOI] [PubMed] [Google Scholar]
- Rasey J. S., Carpenter R. E., Nelson N. J. Response of EMT-6 tumors to single fractions of X rays and cyclotron neutrons. Evaluation and comparison of multiple endpoints. Radiat Res. 1977 Aug;71(2):430–446. [PubMed] [Google Scholar]
- Rockwell S. C., Kallman R. F., Fajardo L. F. Characteristics of a serially transplanted mouse mammary tumor and its tissue-culture-adapted derivative. J Natl Cancer Inst. 1972 Sep;49(3):735–749. [PubMed] [Google Scholar]
- Twentyman P. R. An artefact in clonogenic assays of bleomycin cytotoxicity. Br J Cancer. 1977 Nov;36(5):642–644. doi: 10.1038/bjc.1977.243. [DOI] [PMC free article] [PubMed] [Google Scholar]