兔角膜缘干细胞体外壳聚糖共混膜上培养的形态学观察

Morphology of corneal limbal stem cell culture on chitosan scaffold surface in rabbit

目的 探讨壳聚糖作为兔角膜缘干细胞体外培养载体的可行性。方法 应用消化培养法在壳聚糖材料上对兔角膜缘细胞进行原代培养 ,倒置显微镜观察细胞生长情况 ,并在电子显微镜下观察壳聚糖的结构以及第 7d时细胞的形态。结果 壳聚糖呈现三维多孔结构相互连接成网状 ,孔径直径大小约 90 μm ,其上有细胞生长 ,7d左右形成单层次 ,9d后细胞培养开始出现老化现象。结论 壳聚糖上可成功地对兔角膜缘组织进行原代培养 ,可用作组织工程角膜的载体材料。

Objective To study a feasibility of chitosan as a culture vector for rabbit corneal stem cell in vitro. Methods Rabbit limbus was cultured primarily on chitosan by enzymatic digestion culture method.Cultured cells were observed under inverted microscope every day,and the microstructure of fabricated chitosan scaffold was observed with scanning electron microscope at the 7 th day. Results Fabricated chitosan scaffold exhibited a three-dimensional porous structure with 90 μm pores that created an anastomosing network throughout matrix.Cells seeded on chitosan scaffold were allowed to proliferate and form confluent monolayer in day 7,and the cultured cells appeared to be aging at the 9 th day. Conclusion The epithelial cells from limbus contained stem cells that can be cultured on the surface of chitosan scaffold,and chitosan scaffold is a suitable corneal vector for tissue engineering.