应用三维培养技术体外重建角膜组织的初步研究

Primary research about reconstruction of cornea in three-dimensional collagen gel in vitro

目的 探讨三维培养技术体外重建角膜组织的理论基础及临床应用价值。方法 原代培养人角膜上皮、基质及内皮细胞 ,通过胶原凝胶三维培养系统 ,体外重建角膜结构 ,应用裂隙灯检测人工角膜的透明度 ,采用常规病理组织学及透射电镜观察细胞在三维培养系统中生长的特性。结果重建的角膜组织为良好的三层结构 :角膜上皮细胞形成复层结构 ;内皮细胞形成单层结构 ,且排列规则 ;角膜基质细胞呈散在分布。电镜观察 ,细胞在Ⅰ型胶原中存活并合成胶原纤维。结论 利用组织工程技术 ,能够体外重建结构较规则、透明度较高的角膜组织。

Objective To research the reconstruction of cornea in three dimensional culture. Methods Using primary cultured corneal epithelial cells, stromal cells and endothelial cells rebuilt a corneal structure in three dimensional collagen gel reconstruction system. The artificial corneal transparency was examined by a slit lamp microscope. The biological characters of cells were tested by routine pathologic and transmission electron microscopic technology. Results The reconstructed corneal tissue formed three layers. The epithelial cell layer was two or three cell thickness. The endothelial cells formed a single layer. The stromal cells were diffuse in distribution in the collagen gel. The cells had proliferative ability and capability of producing collagen. Conclusion The regular and transparent corneal structure can be rebuilt by tissue engineering technology.