Breast cancer cells from patients hold an important role in antigen production for immunotherapy, drug
testing, and cancer stem cell studies. To date, although many studies have been conducted to develop protocols for the
isolation and culture of breast cancer cells from tumour biopsies, the efficiencies of these protocols remain low. This
study aimed to identify a suitable medium for the isolation and propagation of primary breast cancer cells from breast
tumour biopsies. Breast tumour biopsies were obtained from hospitals after all patients had given their written informed
consent and were cultured according to the expanding tumour method in 3 different media: DMEM/F12
(Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12) supplemented with 10% FBS (Fetal bovine serum) and
1% antibiotic-antimycotic (Medium D); Medium 171 supplemented with 1X MEGS (Mammary Epithelial Growth
Supplement) and 1% antibiotic-antimycotic (Medium M); or a 1:1 mixture of Medium D and Medium M (Medium
DB). The cell culture efficiency was evaluated by several criteria, including the time of cell appearance, cell morphology,
capability of proliferation, cell surface marker expression, ALDH (Aldehyde dehydrogenases) activity, karyotype,
and tumour formation capacity in immune-deficient mice. Notably, primary cancer cells cultured in Medium DB
showed a high expression of breast cancer stem cell surface markers (including CD44+CD24- and CD49f+), low expression
of stromal cell surface markers (CD90), high ALDH activity, an abnormal karyotype, and high tumour formation
capacity in immune-deficient mice. These findings suggested that Medium DB was suitable to support the survival
and proliferation of primary breast cancer cells as well as to enrich breast cancer stem cells.
Key words: Breast cancer cell, breast cancer stem cell, culture medium, primary cancer cell, tumor biopsy.
|
