An enzymatic method is described for disaggregation of viable tumor cells from human solid tumors. The enzymatic cocktail consists of 0.1% collagenase, 0.01% hyaluronidase, and 0.002% deoxyribonuclease. After mechanical mincing of the tumor tissue, tumor specimens are dissociated by incubation in the enzymatic cocktail for 12-18 hours at room temperature. In 17 cases of sarcoma, the mean yield was 5 X 10(6) viable cells per gram tumor tissue. Yield was 1 X 10(7) viable cells per gram tumor tissue in 23 cases of gastrointestinal carcinoma. The viabilities of tumor cell suspensions ranged from 50 to 98%, except for low viabilities in four specimens that were grossly composed almost entirely of necrotic tissue. The dissociation procedure is simple and the viable cell yield is sufficient for applications in studies of human cancer immunobiology.
A murine monoclonal antibody, CHIP, has been prepared against a human pancreatic carcinoma cell line, SHAW. With the use of the avidin-biotin immunoperoxidase technique, the CHIP antibody detected an antigen found in 11 of 20 fixed tissue sections of tumors obtained from patients with pancreatic carcinoma. The antibody also detected the antigen in 25 of 26 colon carcinoma specimens, 4 of 6 gastric carcinoma specimens, and 1 of 2 esophageal adenocarcinoma specimens. The antigen was also found in normal proximal jejunum and colon and in small amounts in pancreatic islets and parathyroid. There was no reactivity with normal pancreatic ductal or acinar cells or with mesenchymal tissues.
The efficacy of heparin (HEP), the heparin analogue hexuronyl hexosaminoglycan sulfate (HHS), and hydrocortisone (HC) was studied in inhibiting the growth of four morphologically distinct pancreatic adenocarcinoma lines (CBP, LHP2, LSP3, and Pour-LVG) in hamsters. Animals were inoculated with LD100 doses of one of the four tumor lines and were randomly allocated to groups of five animals, which received in their drinking water either: HEP (1000 U/ml) alone, HHS (10 mg/ml) alone, HC (0.5 mg/ml) alone, HEP plus HC, HHS plus HC, or no additives (control). Tumors were measured, growth rates calculated, and nonparametric statistical comparisons made among the median growth rates of all of the treatment groups. All four tumors were tested in the rabbit cornea assay for their ability to induce angiogenesis. Extracts of tumors from control animals as well as from animals treated with HHS plus HC were prepared for quantitative testing in vitro by endothelial cell migration assay. All four tumor lines caused angiogenesis as measured in the rabbit cornea assay. A reduction in median tumor growth rates was observed in animals treated with HHS plus HC bearing the CBP, Pour-LVG, and LSP3 tumors. Similarly, in vitro capillary endothelial cell migration was decreased by HHS plus HC treatment in animals bearing CBP, Pour-LVG, and LSP3 tumors. Animals bearing the LHP2 tumor showed no effect of HHS plus HC treatment on tumor growth rate and no effect on endothelial cell migration. HEP alone, HHS alone, HC alone, and HEP plus HC showed no effect on tumor growth rate in any of the four tumors tested.
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