Breast disease targets and uses thereof
a breast cancer and target technology, applied in the field of molecular markers and therapeutic agents, can solve the problems of high mortality observed in breast cancer patients, and the difficulty of diagnosis and effective treatment of breast cancer
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1. Breast Tissues and Cells
[0360]Breast cancer cell lines were obtained from ATTC. Cancer cell line SK-BR-3 (Cat# HTB-30) was cultured in McCoy's 5a medium supplemented with 10% FBS, 1× penicillin-streptomycin-glutamine, and sodium bicarbonate.
[0361]Non-cancer breast cell line MCF-10A (Cat# CRL-10317) was cultured in MEGM (Mammary Epithelial Growth Medium) supplemented with Singlequots that accompany the media.
[0362]All cells were passaged 1-2 times a week. Cancer cells as well as non-cancer cells were disassociated with 0.05% trypsin. Prior to harvesting for experiments, cells were cultured for 48 hours until 75-80% confluency and disassociated with versene. All tissue culture reagents were purchased from BioFluids.
[0363]Tissue, both normal and tumor samples, are crudely minced and incubated for 60 minutes with periodic agitation at 37° C. in a buffered enzyme cocktail containing Collagenase and Hyaluronidase. Following digestion, the suspension is further treated ...
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