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.
ABSTRACT
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.
Subject(s)
Animals , Mice , Allografts , Biopsy , Follow-Up Studies , Graft Rejection , Kidney , Kidney Transplantation , TransplantsABSTRACT
Objective Both kidneys of mice used To investigate the surgical procedures for establishment of murine kidney transplantation models by using bilaterial donor kidneys as grafts for two recipients.Method Using Balb/c male mice as donors/receptors,through careful dissection and trimming of donor renal vessels and abdominal great vessels,both right and left kidneys were harvested and separately transplanted into two recipients.The anastomosis between the aorta and vena cava of the graft and recipient mouse was made in end to-side manner using the 10-0 sutures.The artery and vein was anastomosed by dual point 180 continuous sutures.The technique of pulling the end of graft ureter into recipient's bladder and external fixation method were used for urethral reconstruction.Five days later the recipient's kidney was removed after transplantation.Result The kidneys in 30 mice were transplanted (15 left kidneys and 15 right kidneys) and model survival rate was 76.7％ (12/15 for the left and 11/15 for the right).The average operating time of donor was 339 min and that of recipient was 46 12 min.Cold ischemia time for the first and second transplantation was about 0.5 h and 1.5 h respectively.Conclusion Through careful microsurgical operation we establish the mouse kidney transplantation models using both donor kidneys as grafts,which reduces the costs of microsurgery experiments.