Claims
- 1. A method of cytogenetic analysis that comprises evaluating DNA of cultured breast cells obtained from aspirate fluid for evidence of chromosomal gains or losses relative to DNA obtained from a normal breast cell in order to determine the presence or absence of premalignant or malignant breast cells, comprising the steps of:(1) aspirating fluid containing cells from a breast of a subject that is to be tested for the presence or absence of premalignant or malignant cells in the breast; (2) placing the samples of fluid obtained in step (1) onto dishes containing conditioned medium composed of (a) supernatant from immortalized mammary epithelial cells and (b) mammary epithelial growth medium; (3) incubating the product of step (2) in a humidified incubator; (4) replenishing the medium in the dishes prepared in step (2) at regular intervals to maintain cell growth; (5) isolating cells from said cultures; (6) preparing DNA from these cultured cells, as well as reference DNA obtained from normal breast cells; (7) amplifying the subject DNA, as well as the referenced DNA; (8) labeling a first aliquot of amplified DNA of the subject cells, and a second aliquot of amplified DNA from the reference normal breast cells; and (9) comparing the labeled DNA of both samples in order to detect chromosomal gains or losses in the subject breast cells, and correlating chromosomal gains or losses to the presence or absence of premalignant or malignant breast cells in the subject.
- 2. A method of claim 1 wherein the growth medium contains, additionally, antibiotics and anti-fungal agents.
- 3. A method of claim 1 wherein the ratio of supernatant from immortalized mammary epithelial cell cultures and from mammary epithelial growth medium is about 1:1.
- 4. A method of claim 1 wherein the incubation temperature is about 37° C.
- 5. A method of claim 1 wherein the method of amplifying DNA is universal amplification.
- 6. A method of claim 1 wherein the gains and losses in chromosomal make-up is evaluated using two different fluorochromes.
Parent Case Info
This application claims the benefit of Provisional Patent Application Ser. No. 60,062,775 filed Oct. 24, 1997.
Non-Patent Literature Citations (5)
Entry |
Kelsey et al. Epidemiology and prevention of breast cancer. Annu. Rev. Public Health vol. 17 pp. 47-67, 1966.* |
Gray et al. Molecular cytogenetics of human breast cancer. Cold Spring Symp. Quant. Biol. vol. 59 pages 645-652, 1994.* |
Telenius et al. Degenerate oligonucleotide-primed PCR: General amplification of target DNA by a single degenerate primer. Genomics vol. 13 pp. 718-725, 1992.* |
Buehring et al. Growth rates of normal and abnormal human mammary epithelia in cell culture. Cancer Res. vol. 36 pp. 3742-3747, 1976.* |
Bano et al. Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines. Biochemistry vol. 31 pp. 610-616, 1992. |
Provisional Applications (1)
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Number |
Date |
Country |
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60/062775 |
Oct 1997 |
US |