Panc 1 crl 1469

The PANC1 (CRL-1469) and MIAPaCa2 (CRL-1420) cell lines were obtained from the American Type Culture Collection. PHsPDAC cells were a gift from Yangchun Xie and they were generated from patient tumor specimens^{70 (link),71 (link)}. WT and Nupr1^−/− mPDAC cells were generated from Nupr1^+/+;Pdx1-cre;LSL-Kras^G12D or Nupr1^−/−;Pdx1-cre;LSL-Kras^G12D mice, respectively, which was a gift from Juan Iovanna (Centre de Recherche en Cancérologie de Marseille, INSERM, France). WT and Nupr1^−/− mPDAC were used at <10 passages. These cells were cultured in Dulbecco’s modified Eagle’s medium (Thermo Fisher Scientific, 11995073) supplemented with 10% heat-inactivated fetal bovine serum (Thermo Fisher Scientific, A3840001) and 1% penicillin and streptomycin (Thermo Fisher Scientific, 15070-063) at 37 °C, 95% humidity, and 5% CO₂. Cell line identity was validated by short tandem repeat profiling, and routine mycoplasma testing was negative for contamination.

The PANC1 (CRL-1469), Capan2 (HTB-80), and MFN1/2^–/– MEFs (CRL-2994) were purchased from the American Type Culture Collection. These cell lines were grown at 37°C, 95% humidity, and 5% CO₂ according to standard mammalian tissue culture protocols and aseptic techniques. In brief, PANC1 and MEFs were cultured in Dulbecco’s Modified Eagle’s Medium (Sigma-Aldrich, D6429), whereas Capan2 was cultured in McCoy’s 5a Medium Modified (Sigma-Aldrich, M4892). All media were supplemented with 10% heat-inactivated (56°C for 30 min) fetal bovine serum (Sigma-Aldrich, F4135) and 5000 units/ml of penicillin and 5000 μg/ml of streptomycin (Thermo Fisher Scientific, 15070063). The identity of the cell line was verified by short tandem repeat analysis, and routine mycoplasma testing was negative for contamination.

All animal procedures were carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was officially approved by German law for the Care and Use of Laboratory Animals and authorized by the regional government of Upper Bavaria, Germany (reference 55.2-1-54-2532-217-2015 from 13 April 2016). All surgical procedures were performed under anesthesia, and all efforts were made to minimize suffering.
As previously described, an orthotopic xenograft pancreatic tumor mouse model in 6-week-old immunosuppressed CD-1^® nude mice (Crl: CD1- Foxn1nu, Charles River Laboratories, Sulzfeld, Germany) was established [21 (link),22 (link)]. The two different established human pancreatic carcinoma cell lines, Panc-1 (CRL-1469) and MiaPaCa-2 (CRL-1420), were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). 2.0 × 10⁶ cells of the Panc-1 or 1.5 × 10⁶ cells of the MiaPaCa-2 cell line were injected into the parenchyma of the pancreas. When tumors reached a median tumor volume of 90 mm³ after a tumor growth of three (MiaPaCa-2) to eight (Panc-1) weeks, treatment planning and irradiation were performed.

The human gastric cancer cell lines (AGS (ECACC ID 89090402) and HGC-27 (ECACC ID 94042256)) were purchased from ECACC (European Collection of Animal Cell Cultures, Salisbury, England, UK). Human pancreatic cancer cell lines (PANC-1 (CRL-1469) and MIA PaCa-2 (CRM-CRL-1420)) were obtained from the American Type Culture Collection (ATCC/LGC Standard, Teddington, England, UK). Human immortalized mesothelial cells, MET5A (CRL-9444) was also obtained from ATCC. Cells were cultured in a humidified incubator at 37 °C, 5% CO₂ and 95% humidity in Dulbecco’s Modified Eagle Medium (DMEM)/F12 (Sigma-Aldrich, Dorset, England, UK) supplemented with 10% fetal Calf Serum (FCS) (Sigma-Aldrich, Dorset, UK) with 1% antibiotics (Sigma-Aldrich, Dorset, England, UK).