Dysfunction of endogenous DCs and tumor-specific T cells in tumor-bearing patients raises the need to develop ex vivo tools to generate therapeutic T cells. Here, we tested the feasibility of generating high numbers of antigen-specific CTLs from peripheral blood lymphocytes (PBLs) for adoptive cell therapies using in vitro sensitization (IVS) with Ag-loaded alpha-type-1 polarized dendritic cells (αDC1), monocyte-derived DCs matured in TNFα/IL-1β/poly-I:C/IFNα/IFNγ. Compared with “standard” DC (sDC; matured in TNFα/IL-1β/IL-6/PGE2), αDC1 induced higher numbers of tumor-specific CTLs, with more diverse T‑cell receptor repertoires and enhanced tumor recognition and killing capacity, without undergoing exhaustion. αDC1‑sensitized tumor-specific CTLs could be enriched based on their 4‑1BB expression and subsequently expanded using rapid expansion protocol (REP), while retaining potent antigen‑specific cytotoxic function. Our study identifies an optimized versatile platform for efficient manufacturing of patient-customized multi-epitope-specific T cells for adoptive cell therapy of heterogenous solid tumors.