肾纤维囊作为载体培养角膜基质细胞的研究

Kidney fiber capsule as a carrier of corneal stromal cell culture

目的探索以脱细胞的肾纤维囊作为载体培养角膜基质细胞并研究培养角膜基质细胞特性。方法将培养扩增的兔角膜基质细胞分别接种到培养板和肾纤维囊上进行体外培养,采用倒置显微镜、免疫组化(波形蛋白)和免疫荧光(AO、PI和Hoechst)的方法进行检测,观察兔角膜基质细胞在培养板和肾纤维囊上的生长情况。结果倒置显微镜和免疫荧光显微镜观察结果显示角膜基质细胞在肾纤维囊上快速贴壁生长并增殖,细胞形态呈多角形树枝状,多次传代后细胞仍维持原有的形态和功能,细胞能长期培养。在培养板上培养的细胞呈长梭形,细胞贴壁生长和增殖情况稍差。在肾纤维囊上的活细胞较在培养板上培养的细胞多。结论角膜基质细胞在脱细胞的肾纤维囊载体上生长良好,细胞形态结构明显。本研究为角膜基质细胞的体外培养,提供了简单和高效的方法。

Objective To investigate the feasibility of kidney fiber capsule as carrier for cultivating corneal stroreal cells and to study the characteristics of the cultured cells. Methods Rabbit corneal stromal cells were seeded on culture plates or kidney fiber capsule in vitro. Stromal cells were identified by observing live cells under inverted microscope and fluorescent microscope. The cultured cells were stained with vimentin, acridine orange, propidiumiodide and Hoechst. Results Inverted and fluorescent microscope showed that comeal stromal cells seeded on the kidney fiber capsule grew well and had r,ormal cell morphology. Cultured cells attached firmly on the surface of kidney fiber capsule. Stromal cells showed triangle or dendritic morphology with many intercellular joints which formed networks. Corneal stromal cells seeded on the culture plates showed long spindle. Its growth and proliferation were superior to the cells on the kidney fiber capsule. Conclusion The kidney fiber capsule has good scaffold property. It facilitates the growth of corneal stromal cells and cultured cells show typical characteristics. This study provide a simple and efficient way culturering stromal cells in vitro. The kidney fiber capsule is an ideal biomaterial for layering tissue engineered cornea stroma.