Ssea3 4

MCF10A and MCF10C breast cell lines were derived at the Barbara Ann Karmanos Cancer Institute (Detroit, MI) and maintained in DMEM-F/12 medium containing 5% heat-inactivated horse serum, 10 μg/mL insulin, 20 ng/mL epidermal growth factor, 0.1 μg/mL cholera enterotoxin, and 0.5 μg/mL hydrocortisone [53 (link),54 (link)]. Breast cancer cell lines MCF7, Hs587T, and MDA231 were purchased from ATCC, and were propagated in 10% fetal bovine serum (Invitrogen, Grand Island, NY); Dulbecco’s Modification of Earle’s Media (Cellgro, Herndon, VA); 2 mM L-Glutamine (Invitrogen); 200 U Penicillin/ml; 200 μg Streptomycin/ml (Invitrogen).
Human breast cancer stem cells (BCSC: CD133+, CD44+, SSEA3/4+, Oct4+, Alkaline Phosphatase+, Aldehyde Dehydrogenase+, Telomerase+), pancreatic cancer stem cells (PCSC: CD44⁺, CD133⁺, SSEA3/4⁺, Oct4⁺, Alkaline Phosphatase⁺, Aldehyde Dehydrogenase⁺, Telomerase⁺, and Nestin⁺), and prostate cancer stem cells (PrCSC: CD44⁺, CD133⁺, SSEA3/4⁺, Oct4⁺, alkaline phosphatase⁺, aldehyde dehydrogenase⁺, and telomerase⁺) were purchased from Celprogen (San Pedro, CA), and cultured using specialized media and tissue culture plastic and matrix, to preserve their CSC phenotype, according to the manufacturer’s instructions.