利用人源化小鼠研究组织工程材料的体内免疫原性  

Experimental Study of Immunogenicity of Tissue Engineering Materials by Using Humanized NOD/SCID Mice

作　　者：张秋玉[1,2] 盛慧明[2] 崔磊[3] 王瑛[2] 尹烁[3] 林锦骠[2] 吴娟娟[2] 沈佰华[2] 王利[2] 李宁丽[2] 

机构地区：[1]福建医科大学免疫系,福建省福州市350004 [2]上海交通大学医学院,上海市免疫学研究所,上海市200025 [3]上海组织工程研究与开发中心,上海市200235

出　　处：《组织工程与重建外科杂志》2009年第4期190-194,共5页Journal of Tissue Engineering and Reconstructive Surgery

摘　　要：目的建立并利用人源化小鼠模型,研究组织工程材料的体内免疫原性。方法利用NOD/SCID小鼠(非肥胖型糖尿病鼠与重症联合免疫缺陷鼠反交鼠模型),通过腹腔注射人外周血单个核细胞(PBMC)(100×106)建立人源化小鼠,在人源化小鼠皮下植入异体人皮肤(阳性对照)、人体脂肪干细胞与脱钙骨构建的组织工程骨(实验组)、单纯脱钙骨材料(阴性对照)。在植入后1周、2周、3周、4周,取小鼠尾静脉血,流式细胞仪检测人CD4+和CD8+淋巴细胞比例。4周后,取小鼠的移植局部皮肤进行HE染色分析,同时检测脾细胞人CD4+和CD8+淋巴细胞比例。结果NOD/SCID小鼠腹腔注射人PBMC4周内,在外周血和脾中均可测到人CD4+和CD8+淋巴细胞。病理分析结果显示:在人源化小鼠皮下植入的组织工程材料及成体干细胞构建的组织工程骨组未见炎性细胞浸润;而单独植入人皮肤组可见明显的炎性细胞浸润。结论人源化小鼠可作为组织工程材料体内免疫原性的研究的良好模型。Objective To investigate the immunogenicity of new transplanted materials in vivo by using humanized NOD/SCID mice model. Methods humanized mice model was established by transferred 100×10^6 human peripheral blood mononuclear cells （PBMC） into NOD/SCID mice. Next day, the mice were received the allotransplantation of pieces of human skin from plastic surgery patients as positive control group and received tissue engineering materials （silica gel） accompanying with human adipose-derived stem cells （ADSCs） as experimental group. Negative control group was transplanted with silica gel alone. The percentage of human CD4^＋ and CD8^＋ T cells in peripheral blood and spleen was detected by fluo-rescent flow cytometry （FACS） at 1, 2, 3 and 4 weeks after operation. Human tymphocytes infiltration within the engrafted section was examined by hematoxylin-eosin staining 4 weeks after transplantation. Results Human CD4^＋ and CD8^＋T cells have been detected both in peripheral blood and spleen of humanized NOD/SCID mice. The percentage of human CD4^＋ and CD8^＋ T ceils in peripheral blood decreased gradually. However, in the mice received the grafts, the percentage of human PBMC decreased much faster. The hematoxylin-eosin staining showed that in the experimental group, wound inflammatory response was slight, without lymphocytes infiltration. In contrast, all human skin grafts were rejected 4 weeks after subcutaneou transplantation. Conclusion These results indicated that the humanized NOD/SCID mice model could be used to analyze the immunogenicity of new transplnted tissue material in vivo.

关 键 词：非肥胖型糖尿病鼠与重症联合免疫缺陷鼠反交鼠模型小鼠 免疫原性 组织工程材料 

分 类 号：R392.12[医药卫生—免疫学]