Characterization of human colon carcinoma cell lines isolated from a single primary tumour

Abstract

Summary The initiation of a cultured human colon carcinoma line on a feeder layer of confluent fibroblasts is described. Attempts to initiate cultures without fibroblast feeder layers were not successful. Two sub-lines (designated HCT C and HCT C Col) were isolated and weaned from cells growing on the surface of the feeder layer. The sub-lines had different morphologies, secreted different levels of carcinoembryonic antigen (CEA) into the medium of confluent cultures and had slightly different karyotypes. Both sub-lines grew in semi-solid medium and formed xenografts when injected s.c. in to athymic nude mice. Analysis of radioiodinated cell membrane components indicated small, but significant differences between the sub-lines.Feeder layers of normal cells have frequently been utilized as an aid in the establishment of tissue cultures from a variety of human tumour specimens including mammary carcinoma (Smith et al., 1981), lymphoma (Epstein and Kaplan, 1979) and retinoblastoma (Gallie et al., 1982). Recently, we described the establishment of human colonic carcinomas in tissue culture (Brattain et al., 1981a). We observed that some primary cultures contained several types of malignant cells as judged by the different morphologies of cells growing on the surface of the fibroblast feeder layer. Heterogeneity of the malignant cell compartment comprising human solid tumours is currently recognized as a potentially important obstacle in several therapeutic treatment modalities of cancer Heppner et al., 1978). In addition to the clinical problems posed by heterogeneity of the malignant cells, minority subpopulations may contribute significantly to the determination of such biological properties of the tumour as metastatic or invasive abilities (Fidler, 1978;Poste & Fidler, 1980). Consequently, the identification and characterization of subpopulations of malignant cells from individual tumours is of potential importance to both the treatment and understanding of cancer. Utilizing a feeder layer of normal human colon fibroblasts we have identified two morphological subpopulations of cells from primary tissue cultures of a human colon tumour. We describe the establishment, isolation and characterization of two sub-lines of malignant cells from these primary cultures. Materials and methodsTissue culture A human colon carcinoma (designated HCT C) and normal human colon were provided by the UAB Tissue Procurement Service. The sample of normal colon was obtained at the time of colon resection for carcinoma in February, 1979 while the resection for HCT C was performed in April, 1979 on a different patient. Procedures for preparing the specimens for culture were identical and have been described for colon carcinomas in detail (Brattain et al., 1977a, b;1979;Brattain, 1979). Briefly, the specimens were minced to small pieces (1-2 mm3) and extensively washed in McCoy's tissue culture medium supplement with 20% fetal bovine serum (FBS) and antibiotics (4.3 /ugml-P gentamicin, 90 jugml-l streptomycin, 90 Uml-penicillin and 2.5...

Role of cancer-associated stromal fibroblasts in metastatic colon cancer to the liver and their expression profiles

“…To characterize the features of the CAFs, we showed by conditioned-medium proliferation assays that CAF-derived medium promoted cancer cell proliferation to a greater extent than other fibroblastderived medium, likely by secreting more growthpromoting factors (Gleave et al, 1991). Irrespective of the origin of fibroblasts, previous reports have shown that fibroblasts enhance the growth of cancer cells and are often required to establish tumor by providing a feeding layer (Brattain et al, 1982;Picard et al, 1986;Camps et al, 1990;Gleave et al, 1991). In our conditioned-medium assays, without direct interaction between cancer cells and fibroblasts, we also observed a clear-cut enhancement of cancer cell growth by all fibroblasts.…”

Orthotopic implantation of human colon carcinomas into nude mice provides a valuable model for the biology and therapy of metastasis

“…Xenografts of colon tumors in nude mice have been produced either directly from surgical specimens [18,19,[24][25][26][27] or from cell lines established in culture [25,[27][28][29][30]. Whether fresh specimens or cell lines provide the better model for HCC in nude mice is still debated.…”

Stability of critical genetic lesions in human colorectal carcinoma xenografts

“…However, for none of the long-established colorectal cancer lines readily available is status at critical oncosuppressor loci known. Moreover long-established lines commonly acquire additional genetic rearrangements during culture (Brattain et al, 1983;Park et al, 1987), which may not be reversible by correction of the original oncosuppressor defect alone. We therefore set out to develop a series of lines from primary colorectal tumours, defined both in terms of status at oncosuppressor loci on chromosomes 5, 17 and 18, and the stability of each locus on serial passage.…”