组织工程皮肤细胞对朗格汉斯细胞表型的影响

Effect of engineered heterogeneous skins cells on phenotypes of langerhans cells

目的:研究异体组织工程皮肤种子细胞(角质形成细胞、成纤维细胞)对朗格汉斯细胞的表型影响。方法:外周血单个核细胞在粒细胞巨噬细胞集落刺激因子(GM-CSF)、白介素4(IL-4)和转化生长因子β1(TGF-β1)的诱导下培养出朗格汉斯细胞。培养的朗格汉斯细胞与异体组织工程皮肤种子细胞共同培养后,检测其表型变化情况。随后将朗格汉斯细胞与淋巴细胞共培养后检测淋巴细胞的增殖程度。结果:诱导培养的朗格汉斯细胞低表达HLA-DR、CD80和CD86,不表达CD83。与异体组织工程皮肤种子细胞共培养后朗格汉斯细胞表型无明显变化,共培养后的朗格汉斯细胞无法刺激自体淋巴细胞增殖。结论:与异体组织工程皮肤种子细胞共培养后,朗格汉斯细胞仍保持不成熟的状态,这提示组织工程皮肤种子细胞免疫原性较低,移植后不易引起宿主的免疫排斥反应。

Objective： To study the effect of seeding cells of tissue-engineered skins, keratinocytes and fibroblasts, on the phenotypes of langerhans cells （LC）. Methods： Peripheral blood mononuclear cells were induced by cytokines, granulocyte macrophage-colony stimulating factor （GM-CSF）, interleukin 4 （IL-4）, transforming growth factor β1 （TGF-β1） to generate LCs, which were subsequently cocultured with allogenic keratinocytes or fibroblasts. Then the phenotypes of langerhans cells was detected by flow cytometer （FCM）, and the degree of proliferation of autogeneic lymphocytes was assessed after stimulation with LCs. Results： Cultured LCs had low level expression of HLA-DR, CDS0, CD86 and no expression of CD83. Cocultured with allogenic keratinocytes or fibreblasts, phenotypes of LCs did not change significantly. Simultaneously, LCs could not stimulate autogeneic lymphocytes proliferation. Conclusion： These data indicate that LCs cocultured with seeding cells of tissue-engineered skins remain immature state. It is suggested that seeding cells of tissue-engineered skins have low immunogenicity and are unable to induce significant immunological rejection of host after transplantation.