Generation of in vitro porcine monocyte-derived dendritic cells (MoDcs) by new modified protocol

Abstract

The derivation of dendritic cells (Des) in vitro is an alternative system to overcome the low frequency of primary DCs and the difficulty of isolation techniques for study of DC immunobiology. To date, conventional culture protocol of porcine monocyte-derived DCs (MoDCs) has been widely used. However, this protocol is sometimes not practical due to the requirement of substantial monocyte number from the blood sample, and the process often interferes with DC maturation. To improve the protocol for porcine MoDC generation, we altered the previous conventional protocol, based on the human MoDC and mouse bone-marrow derived DC (8M-DC) culture system, and compared phenotypic and functional features of MoDC derived from the modified protocol to the conventional protocol. The modified protocol consumed less amount of monocytes but generated higher CD1 + cells with DC-like morphology and ability of maturation. In addition, MoDCs from the modified protocol exhibited increased antigen uptake capability and cytokine gene expression in response to LPS stimulation. Our findings indicate that the modified protocol is expedient and reliable for generating potent MoDCs that substitute for primary DCs. This will be a valuable platform for future research in antigen delivery, vaccine and immunotherapy in pigs, as well as relevant veterinary species.