Mus musculus
Protocols: (Mouse organoids from Bag6 WT, Bag6 KO and PBS control groups were collected in Eppendorf tubes (n=3 per each group). Organoids were dissociated in cold PBS to remove the Matrigel. Cells were washed twice with cold PBS and centrifuged at 300 xg for 5 minutes at 4°C. Pancreatic cancer organoids were derived from the KPC mouse model according to published procedures (1). The KPC organoids (5,000 cells in 20 µL matrigel per 96 well) were cultured in organoid growth medium (1) plus the secretome (ratio 1:1) derived from mouse mast cells pre-treated according to the following conditions: 1. PBS: Mast cells pre-treated with PBS (control). 2. KO-EVs: Mast cells pre-treated with Bag6 KO EVs. 3. WT-EVs: Mast cells pre-treated with Bag6 WT EVs. After 5 days, phase contrast images were taken by confocal microscope (Leica DMI3000 B) and cell percentage was assessed using the Cell Titer 3D GLO kit (Promega, Walldorf, Germany), control PBS was set to 100%. RNA was extracted at the same timepoint. References: (1). Boj SF, Hwang CI, Baker LA, et al. Organoid models of human and mouse ductal pancreatic cancer. Cell 2015;160:324-38. Subsequently, the cells were lysed, and total RNA was isolated using the RNeasy Plus kit (Qiagen, Hilden, Germany, [Cat. No. / ID: 74034]) according to the manufacturer's instructions). Lexogen QuantSeq according to Manufacture's Instructions
disease
pancreatic carcinoma
organism
Mus musculus
organism
Mus musculus
collection date
not collected
scientific_name
Mus musculus
common name
house mouse
organism part
pancreatic duct
developmental stage
adult
genotype
Kras+/LSL-G12D; Trp53+/LSL-R172H; Pdx1-Cre
isolate
not applicable
growth condition
conditioned media of WT Mast cells
cell_line
KPC
cell_type
organoid
geographic location (country and/or sea)
not collected
ENA-LAST-UPDATE
2023-11-24