Toxicity to the normal hemocytes by ALA-PDT for the ex vivo purging of hematopoietic stem cell grafts  

作　　者：张宝琴 张镇西 苗丽霞 谭璐 肖谧 徐霞 

机构地区：[1]Department of Pediatrics, the First Affiliated Hospital, Medical School of Xi'an Jiaotong University [2]The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University

出　　处：《Journal of Pharmaceutical Analysis》2008年第4期250-255,共6页药物分析学报（英文版）

基　　金：supported by the National Natural Science Foundation of China (No.60178034);Doctor Thesis Founda-tion of Xi an Jiaotong University (No. DFXJTU2002-2) .

摘　　要：Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control; Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change; however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA, except for PBMCs, intracellular PpIX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosis. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologous bone marrow or stem cell grafts.Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control; Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change; however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA, except for PBMCs, intracellular PpIX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosis. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologous bone marrow or stem cell grafts.

关 键 词：5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) peripheral blood mononuclear cell peripheral blood stem cell cord blood mononuclear cell autologous stem cell transplantation 

分 类 号：R733.7[医药卫生—肿瘤]