Establishment of a Repository for Cell Cultures and Genomic DNA from Breast Cancer Patients.
Final rept. 1 Sep 94-30 Sep 97,
BIOLOGICAL RESEARCH FACULTY AND FACILITY INC IJAMSVILLE MD
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New breast cell lines and cell strains were developed from breast cancer tissues, uninvolved tissues from cancer patients, and normal tissues from reductive mammoplasty patients. Primary cultures were initiated using cell suspensions isolated from tissues and using cellular outgrowth from explanted tissues. Breast cultures were transfected with pRSV-T plasmid or transfected with a plasmid containing mutated ras oncogene or were treated with a mutagenic chemical. Various cell isolation methods and new serum-free media were used for developing cell cultures. Twenty cell strains and 10 cell lines 8 epithelial and 2 fibroblastic were established and added to the breast cell repository. The majority of the epithelial lines have been in continuous culture for at least 20 weeks. The karyotype, cumulative population doubling levels, anchorage-independent growth, and tumorigenicity in nude mice for most of the established cell lines have been documented. Most cell lines possess the U-ras, K-ras, and p53 genes and express the cytokeratin 8 and 18, BRCA-l, erbB2, and EGFR genes. Two anticancer agents, taxol and 5-fluorouracil, produced significant cytotoxic effects on one of the new cancer cell lines. Genomic DNA was isolated from the epithelial lines and stored in the DNA bank.
- Anatomy and Physiology
- Medicine and Medical Research