Generation and Characterization of Cell Lines for CAR-T Therapy

Murine Panc02-EpCAM, 4T1, T110299 and CT26-EpCAM have been previously described [41 (link)–43 ]. Murine LL/2 were purchased from the European Collection of Authenticated Cell Cultures (ECACC). The T110299 and LL/2 cell lines were modified to stably express full-length murine EpCAM (UNIPROT entry Q99JW5), to generate the cell lines T110299-EpCAM and LL/2-EpCAM. Human SUIT-2-MSLN have been previously described [41 (link)]. 293Vec-Galv, 293Vec-Eco, and 293Vec-RD114 have been previously described [44 (link)]. The virus producing cell lines 293Vec-Eco for anti-EpCAM-CAR-28z and 293Vec-RD114 for anti-MSLN-CAR-28z have been previously described [45 (link)]. 293Vec-RD114 for anti-MSLN-CAR-4-1BBz were generated as previously described [46 (link)]. All cell lines were cultured as previously described [41 (link), 42 (link)]. All cell lines were periodically tested for mycoplasma contamination with the commercial testing kit MycoAlert^TM (Lonza, Basel, Switzerland). Authentication of human cell lines by short tandem repeat DNA profiling was conducted in-house.