Triple negative breast cancer immune cells were processed as previously described [12 (link)]. Briefly, fresh TNBC core biopsy tumor specimens were received in MACS Tissue Storage Solution (Cat #130-100-008) on ice and minced in 10 cm ultra-low attachment dishes (Cat # 3262, Corning), and digested with Rat tail collagenase IV (Cat # 17104019, Life Technologies) for 30 min. Red blood cells were lysed with ACK buffer (Cat # 420301, BioLegend) and single cells were obtained and stained with DAPI (Cat # 422801, Biolegend) and Calcein (Cat # 425201, Biolegend). The BD Aria III sorter was used with a 100 μm nozzle to sort live (DAPI- Calcein+) single cells.
Protocol detail
Isolation of TNBC Immune Cells
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Aria
Buffer
Calcein
Cells
Collagenase
Core biopsy
Dapi
Dishes
Red blood cells
Tail
Tissue
Triple negative breast cancer
Tumor
Corresponding Organization :
Other organizations : NYU Langone Health, University of North Carolina at Chapel Hill, New York Genome Center, New York University
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Variable analysis
independent variables
- Mincing TNBC core biopsy tumor specimens in 10 cm ultra-low attachment dishes
- Digesting with Rat tail collagenase IV for 30 min
- Live (DAPI- Calcein+) single cells
- MACS Tissue Storage Solution (Cat #130-100-008)
- ACK buffer (Cat # 420301, BioLegend) for red blood cell lysis
- DAPI (Cat # 422801, Biolegend) and Calcein (Cat # 425201, Biolegend) staining
- BD Aria III sorter with a 100 μm nozzle
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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