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Culture and identification of regulatory dendritic cells from murine bone marrow induced by 5-Aminolevulinic acid combined with ferrous iron in vitro / 中华器官移植杂志
Chinese Journal of Organ Transplantation ; (12): 627-631, 2018.
Article in Chinese | WPRIM | ID: wpr-734830
ABSTRACT
Objective To explore a method to generate regulatory dendritic cells (DCregs) from murine bone marrow induced by 5-Aminolevulinic acid combined with ferrous iron (ALA/SFC).Methods Bone marrow cells were obtained from male C57BL/6 mice.To generate conventional DCs (BM-DCcons),the ceils were cultured in RPMI-1640 medium supplemented with 10% FCS,10 ng/mL GM-CSF,10 ng/mL IL-4 for 7 days.The cells were collected for the analysis.To generate DCregs by ALA/SFC,the cells were cultured in RPMI-1640 medium supplemented with 10% FCS,20 ng/mL GM-CSF,5-ALA 1 mmol/L + SFC 0.5 mmol/L for 7 days.The morphology of ALA-DCregs was observed by microscope and eytospin with May-Grunwal&Giemsa stain.The surface markers of ALA-DCregs were observed by FACS.The function of ALA-DCregs was detected by in vitro mixed lymphocyte reaction (MLR) and in vivo lymphocyte proliferation assay.Results The generated ALA-DCregs displayed an irregular shape with areas of protrusion and demonstrated higher CD11b/CD11c and higher MHC-II but lower CD40,CD80,CD86 expression levels than DCcons.They also had immune regulation effects in both in vitro and in vivo lymphocyte proliferation assay.Conclusion This study illustrated a feasible approach for generating functional DCregs from murine bone marrow induced by ALA/SFC.These cells can be useful for research and application of DC immunotherapy in the future.

Full text: Available Index: WPRIM (Western Pacific) Type of study: Diagnostic study Language: Chinese Journal: Chinese Journal of Organ Transplantation Year: 2018 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Type of study: Diagnostic study Language: Chinese Journal: Chinese Journal of Organ Transplantation Year: 2018 Type: Article