title: Generation of potent porcine monocyte-derived dendritic cells (MoDCs) by modified culture protocol.
abstract: In vitro derivation of dendritic cells (DCs) is an alternative approach to overcome the low frequency of primary DCs and the difficulty of isolation techniques for studying DC immunobiology. To date, the conventional culture protocol of porcine monocyte-derived DCs (MoDCs) has been widely used. However, this protocol is not practical due to the requirement of a substantial number of blood monocytes, and the process often interferes with DC maturation. To improve in vitro porcine MoDC generation, we modified the previous conventional DC generation protocol, based on the human and mouse primary DC culture system, and compared phenotypic and functional features of MoDCs derived from the modified protocol to the conventional protocol. The modified protocol consumed fewer monocytes but generated higher CD1+cells with DC-like morphology and the ability of maturation. In addition, MoDCs from the modified protocol exhibited increased antigen uptake and IFN-γ production 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, vaccines and immunotherapy in pigs, as well as relevant veterinary species.
authors.affiliation: Inter-department of Medical Microbiology, Graduate School, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
authors.forename: Teerawut
authors.initials: T
authors.lastname: Nedumpun
authors.affiliation: RU in Oral Microbiology and Immunology, Department of Microbiology, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand; Oral Biology Research Center, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand. Electronic address: Patcharee.R@chula.ac.th.
authors.forename: Patcharee
authors.initials: P
authors.lastname: Ritprajak
authors.affiliation: Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand; Center of Excellence in Emerging Infectious Diseases in Animals, Chulalongkorn University (CU-EIDAs), Pathumwan, Bangkok 10330, Thailand.
authors.forename: Sanipa
authors.initials: S
authors.lastname: Suradhat
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D018949:Antigens, CD1 / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D009000:Monocytes / Q000276:immunology; D010587:Phagocytosis; D034421:Sus scrofa / Q000235:genetics / Q000276:immunology*
chemical_list: D018949:Antigens, CD1; D007371:Interferon-gamma
keywords: Conventional protocol; Dendritic cell; In vitro culture; Modified protocol; Porcine monocyte derived-dendritic cell