Drug and radiation sensitivity measurements of successful primary monolayer culturing of human tumor cells using cell-adhesive matrix and supplemented medium
The limitations of the agar suspension culture method for primary culturing of human tumor cells prompted development of a monolayer system optimized for cell adhesion and growth. This method grew 83% of fresh human tumor cell biopsy specimens, cultured and not contaminated, from a heterogeneous group of 396 tumors including lung cancer (93 of 114, 82%); melanoma (54 of 72, 75%); sarcoma (46 of 59, 78%); breast cancer (35 of 39, 90%); ovarian cancer (16 of 21, 76%); and a miscellaneous group consisting of gastrointestinal, genitourinary, mesothelioma, and unknown primaries (78 of 91, 86%). Cell growth was characterized morphologically with Papanicolaoustained coverslip cultures and cytogenetically with Giemsastained metaphase spreads. Morphological features such as nuclear pleomorphism, chromatin condensation, basophilic cytoplasm, and melanin pigmentation were routinely seen. Aneuploid metaphases were seen in 90% of evaluable cultures, with 15 of 28 showing 70% or more aneuploid metaphases. Colony-forming efficiency ranged between 0.01 and 1% of viable tumor cells, with a median efficiency of 0.2%. This culture system uses a low inoculum of 25,000 viable cells per well which permitted chemosensitivity testing of nine drugs at four doses in duplicate from 2.2 X 10(6) viable tumor cells and radiation sensitivity testing at five doses in quadruplicate from 0.6 X 10(6) cells. Cultures were analyzed for survival by computerized image analysis of crystal violet-stained cells. Drug sensitivity studies showed variability in sensitivity and in survival curve shape with exponential cell killing for cisplatin, Adriamycin, and etoposide, and shouldered survival curves for 5-fluorouracil frequently seen. Radiation sensitivity studies also showed variability in both sensitivity and survival curve shape. Many cultures showed exponential cell killing, although others had shouldered survival curves.
- Research Organization:
- Univ. of Texas, Houston
- OSTI ID:
- 5843390
- Journal Information:
- Cancer Res.; (United States), Vol. 3
- Country of Publication:
- United States
- Language:
- English
Similar Records
Radiation response of drug-resistant variants of a human breast cancer cell line
Drug and radiation sensitivity testing of primary human tumor cells using the adhesive-tumor-cell culture system (ATCCS)
Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
TUMOR CELLS
RADIOSENSITIVITY
BIOPSY
CELL CULTURES
CELL CYCLE
CULTURE MEDIA
NEOPLASMS
ANIMAL CELLS
DIAGNOSTIC TECHNIQUES
DISEASES
560121* - Radiation Effects on Cells- External Source- (-1987)
550603 - Medicine- External Radiation in Therapy- (1980-)