Two breast cancer stem cell lines capable of being stably cultured and keeping original characteristics, separation method and uses thereof
A breast cancer stem cell, stable technology, applied in the field of medicine and biology, to achieve the effect of stable and repeatable passage
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[0032] 1. Isolation and culture of clinically derived breast cancer stem cell lines that can be stably passaged and maintain their original characteristics
[0033] (1) Take freshly resected breast cancer tumor specimens, add 250 units of collagenase III per milliliter of tissue fluid, place them in a constant temperature incubator at 37°C for 4 hours for digestion, and then use a 40mm filter to filter;
[0034] (2) CD44 was sorted by flow cytometry (CytomicsFC500) + , CD24 - and ALDH1 + Cells, regulation requires cell purity above 98% (CD44 and CD24 antibodies are from CellSignaling; ALDH1 antibody is from Technologies);
[0035](3), preparation of breast cancer stem cell culture medium: add 10ng / mL basic fibroblast growth factor (purchased from Invitrogen Company), 20ng / mL epidermal growth factor (purchased from Invitrogen Company) to serum-free DMEM-F12 (purchased from Invitrogen Company) Invitrogen), 2% B27 (purchased from Invitrogen Company) and 5ug / mL insulin (purchas...
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