Isolation and characterization of the first Slovenian human triple‐negative breast cancer cell line

MFUM-BrTNBC-1, a Newly Established Patient-Derived Triple-Negative Breast Cancer Cell Line: Molecular Characterisation, Genetic Stability, and Comprehensive Comparison with Commercial Breast Cancer Cell Lines

“…Cell lines: For this study, 4 CLs were used: our own primary CL MFUM-BrTNBC-1 [ 26 ] and 3 commercially available and widely used BC CLs (MCF-7 [ERα + , ERβ + , PR + , HER2 − ], MDA-MB-231 [Erα − , Erβ − , PR − , HER2 − ], and MDA-MB-453 [Erα − , ERβ + , PR − , HER2 + ]). MFUM-BrTNBC-1 was isolated from a 47-year-old female patient who underwent a standard BC surgical procedure at the University Medical Centre Maribor, Slovenia.…”

“…Histological examination revealed an invasive carcinoma of no special type (NST), G3, with a triple-negative receptor status and Ki67 PI of 90%. The isolation procedure was described in detail in our previous study [ 26 ]. Cell isolation was carried out in a laminar flow cabinet under sterile conditions (Laminar Air Flow Chamber, ISKRA-Pio, Šentjernej, Slovenia).…”

“…A base cell isolation protocol commonly applied in our laboratories and optimised to address cell type-specific needs has been described in our previous articles [ 26 , 35 , 36 ]. The multi-step protocol [ 26 ] used for the MFUM-BrTNBC-1 isolation can be briefly summarised as follows: (1) after the arrival of the tissue that had been macroscopically selected from the pathologist, the tissue was transferred into two Petri dishes; (2) it was twice rinsed with PBS, which contained additional penicillin and streptomycin; (3) the tissue was then soaked with PBS; (4) necrotic and adipose tissue were macroscopically removed; (5) the tumour tissue was stored in a special Petri dish containing a mixture of 0.25 wt.% trypsin/EDTA; (6) mechanical homogenisation of the tissue was performed with a scalpel (tissue pieces with an average size of 2 mm 3 ) while being soaked in trypsin to prevent dehydration; (7) a 1 h incubation period followed; (8) next, the suspension was soaked with Advanced DMEM/F12 s 5 wt.% FBS and split into four 50 mL centrifuge tubes; (9) after centrifugation, the supernatant is discarded and the remaining cell sediment was washed two times with medium (2500 rpm, 15 min); (10) the cells were then resuspended in medium mixed with 5 wt.% FBS, transferred into small Petri dishes for incubation and placed into an incubator.…”

“…Growing cells were regularly observed with an Axiovert 40 inverted optical microscope (Zeiss, Oberkochen, Germany) at several magnifications. As briefly (not in-depth) discussed in our previous publication [ 26 ], the CL was cultured using 2 different culturing procedures. The difference between the procedures was either allowing for a spontaneous overgrowth of the cancer cells or manual sub-selection of epithelial cells from the beginning, immediately after obtaining the primary cell culture.…”

“…Bearing this in mind, we believe that there is a need for a new generation of BC CLs—CLs that are appropriately documented and thoroughly characterised according to international standards from the very beginning (see Supplementary document, Table S1 ) [ 23 , 24 , 25 ]. This research study aimed to comprehensively describe our patient-derived human TNBC-CL (MFUM-BrTNBC-1) and evaluate it in comparison with commercial BC CLs to obtain a well-defined characterisation (in accordance with international CL cultivation standards), allowing for its efficient use in further TNBC-related studies [ 26 ].…”