兔子宫内膜片层体外构建的实验研究

Reconstruction of Tissue-engineered Endometrium Sheet in Vitro

探索以扩增培养的子宫内膜细胞为种子细胞、以液态胶原为支架材料,构建组织工程化子宫内膜片层的可行性。体外分离并规模化扩增子宫内膜上皮细胞与基质细胞,以Ⅰ型液态鼠尾胶原为支架,按自然子宫内膜的结构在体外重建具有两层结构的组织工程化子宫内膜,体外培养14天时进行H.E.染色与免疫组化鉴定。H.E.染色与免疫组化染色结果表明:再造子宫内膜片层在胶原材料中能够较好地维持双层结构,上皮层呈CK18阳性,在某些部位还能够形成极化的柱状上皮。利用体外扩增培养的子宫内膜上皮细胞与基质细胞构建的组织工程化子宫内膜片层具有子宫内膜组织的双层结构与极化上皮特征。

To reconstruct tissue - engineered endometrium sheet by using proliferated endometrial cells and hquid collagen. Rabbit endometrial epithehal cells and stromal cells were isolated and proliferated in vitro. For the reconstruction of tissue engineered endometrium, endometrial stromal cells were mixed with hquid type - I collagen to form cell - collagen construct. After the construct geled, the epithehal cells were inoculated on top of the construct, After 14 days of in vitro culture, H.E. staining and immunohistochemical staining were used to analyse engineered endometrium sheet. H.E. showed that the reconstructed endometrium sheet formed two layered structure. Cells in epithelial layer were positively immuno- stained by anti - cytokeratin antibody and some epithelial cells showed polarized morphology which was more similar to normal endometrial cells. It proves that tissue engineered endometrium sheet reconstructed with proliferated endometrial cells and type I collagen possesses the two- layer structure and polarized epithehal character which is similar to native endometrium.