Generation of cDC-like cells from human induced pluripotent stem cells via Notch signaling [RNA-Seq]

Dendritic cells (DCs) play critical roles in regulating the innate and adaptive immune responses, and have long been a major focus of cancer immunotherapy. Accumulating evidence suggests that conventional type 1 DC (cDC1s) excel in cross-presentation of exogenous antigens on MHC-I molecules, and induction of antitumor CD8+ T cell immunity; however, obtaining large numbers of cDC1s is difficult. The use of reprogramming and differentiation technology is advantageous for obtaining unlimited numbers of autologous cDC1s especially for therapeutic interventions where repeated vaccinations are required. However, generation of cDC1s from human induced pluripotent stem cells (iPSCs) remains elusive. Overall design: iPSC-derived DCs and PBMCs were stained with a master mix of antibodies for surface staining including Live/Dead Fixable Aqua Dead, XCR1, CLEC9A, CD11c and HLA-DR after Fc block with human IgG. CD11c+HLA-DR+CLEC9A+XCR1+ cells were sorted on BD FACSAria II. Total RNA was prepared from these sorted DCs using the miRNeasy mini kit (Qiagen).