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

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.

Noninvasive monitoring of mouse renal allograft rejection using micro-CT

PURPOSE: Acute renal graft rejection can only be definitively diagnosed by renal biopsy. However, biopsies carry a risk of renal transplant injury and loss. Micro-CT is widely used in preclinical studies of small animals. Here, we propose micro-CT could noninvasively monitor and evaluate renal location and function in a mouse kidney transplant model. METHODS: Orthotopic kidney transplantation was performed in a BALB/c -to- C57BL/6j or C57BL/6j-to- C57BL/6j mouse model. After optimizing imaging techniques, five mice were imaged with micro-CT and the findings were verified histologically. RESULTS: Micro-CT can monitor and evaluate renal location and function after orthotopic kidney transplantation. There were no mice deaths while renal transplants were failure. CONCLUSION: We propose that graft micro-CT imaging is a new option that is noninvasive and specific, and can aid in early detection and follow-up of acute renal rejection. This method is potentially useful to improve posttransplant rejection monitoring.