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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 establish and use a humanized mouse model to study the in vivo immunogenicity of tissue engineered materials. Methods Humanized mice were established by intraperitoneal injection of human peripheral blood mononuclear cells (PBMCs) (100 × 106) in NOD / SCID mice (non-obese diabetic mice and severe combined immunodeficiency mouse anti-injection model) Humanized mice were subcutaneously implanted with allogeneic human skin (positive control), tissue engineered bone constructed with human adipose-derived stem cells and decalcified bone (experimental group), and pure decalcified bone material (negative control). After 1 week, 2 weeks, 3 weeks and 4 weeks after implantation, the tail vein blood was collected and the proportions of human CD4 + and CD8 + lymphocytes were measured by flow cytometry. After 4 weeks, the transplanted skin of the mice was taken for HE staining, and the proportion of human CD4 + and CD8 + lymphocytes in the spleen cells was also detected. Results Four weeks after intraperitoneal injection of PBMC in NOD / SCID mice, both human CD4 + and CD8 + lymphocytes were detected in peripheral blood and spleen. Pathological analysis showed that there was no inflammatory cell infiltration in the tissue engineered bone implanted subcutaneously in humanized mice and in adult stem cells. However, inflammatory cell infiltration was observed in human skin implanted alone. Conclusion Humanized mice can be used as a good model for studying the immunogenicity of tissue engineering materials in vivo.