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Competent antigen-presenting cells are generated from the long-term culture of splenocytes with granulocyte-macrophage colony-stimulating factor

 Seul Hye Ryu  ;  Hye Young Na  ;  Moah Sohn  ;  Wanho Choi  ;  Hyunju In  ;  Hyun Soo Shin  ;  Jae-Hoon Choi  ;  Chae Gyu Park 
 IMMUNOLOGY LETTERS, Vol.188 : 96-107, 2017 
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Animals ; Antigen Presentation/immunology ; Antigen-Presenting Cells/cytology ; Antigen-Presenting Cells/drug effects* ; Antigen-Presenting Cells/immunology* ; Antigen-Presenting Cells/metabolism ; Biomarkers ; Bone Marrow ; Cells ; Cell Culture Techniques ; Cells, Cultured ; Dendritic Cells/immunology ; Dendritic Cells/metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology* ; Histocompatibility Antigens Class II/immunology ; Immunophenotyping ; Membrane Proteins/metabolism ; Mice ; Mice, Transgenic ; Signal Transduction ; Spleen/cytology* ; Spleen/immunology* ; Spleen/metabolism ; T-Lymphocyte Subsets/immunology ; T-Lymphocyte Subsets/metabolism ; fms-Like Tyrosine Kinase 3/metabolism
Antigen-presenting cells ; Dendritic cells ; GM-CSF ; Hematopoiesis ; Spleen
Dendritic cells (DCs) are routinely produced from the culture of mouse bone marrow (BM) with granulocyte-macrophage colony-stimulating factor (GM-CSF) within a period of 10days. Although splenic extramedullary myelopoiesis was suggested to occur under the influence of GM-CSF, the hematopoietic outcome of splenic culture with GM-CSF has not been scrutinized. We have cultured mouse splenocytes with GM-CSF for an extended period of time, where we discovered that the CD11b⁺CD11c⁺ cells began to proliferate prominently after 10days and their number increased until the 4th week of the culture. In parallel experiments, FMS-like tyrosine kinase 3 (FLT3) and its ligand, FLT3L, were not found to influence the culture of splenocytes. Like DCs in the culture of BM with GM-CSF, a distinct population of CD11b⁺CD11c⁺MHC IIhi cells was readily identified as DCs in the long-term culture of splenocytes. After being isolated and plated overnight the CD11b⁺CD11c⁺MHC IIhi cells exhibited non-adherent dendritic morphology, while the other CD11b⁺CD11c⁺ cells became adherent. Besides, these CD11b⁺CD11c⁺MHC IIhi cells possessed relatively weak endocytic and phagocytic abilities but displayed strong antigen-presenting capacities, revealing DC-like characteristics; in contrast, the other CD11b⁺CD11c⁺ cells showed strong endocytosis and phagocytosis of antigens but were poor at antigen presentation, indicating macrophage-like traits. Therefore, we demonstrated that phenotypically as well as functionally genuine DCs are generated in the long-term culture of splenocytes with GM-CSF.
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1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
Yonsei Authors
Na, Hye Young(나혜영) ORCID logo https://orcid.org/0000-0002-2886-9926
Park, Chae Gyu(박채규) ORCID logo https://orcid.org/0000-0003-1906-1308
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