Culture and identification of regulatory dendritic cells from murine bone marrow induced by 5-Aminolevulinic acid combined with ferrous iron in vitro
10.3760/cma.j.issn.0254-1785.2018.09.012
- VernacularTitle:5-酮戊二酸(ALA)联合二价铁离子体外诱导小鼠骨髓来源调节性树突状细胞的培养及鉴定
- Author:
Qi ZHANG
1
,
2
;
Songjie CAI
;
Yating LIU
;
Jiangang HOU
Author Information
1. 200040上海,复旦大学附属华山医院皮肤科
2. 200040 上海市皮肤病研究所
- Keywords:
5-aminolevulinic acid;
Dendritic cells;
Mouse
- From:
Chinese Journal of Organ Transplantation
2018;39(10):627-631
- CountryChina
- Language:Chinese
-
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.
