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目的 探讨细胞外基质对淋巴管新生的作用。 方法 在Ⅰ型胶原蛋白凝胶中和Matrigel基膜基质凝胶上培养狗胸导管内皮细胞 ,相差倒置显微镜下观察内皮细胞形成的管状结构的形态 ,透射电镜下观察淋巴管样结构 ,用图像分析仪对管状结构作定量分析。 结果 淋巴管内皮细胞在Ⅰ型胶原蛋白凝胶上生长成单层 ,但未见管状结构形成。在细胞上面覆盖上层凝胶后 ,内皮细胞发生形态变化 ,形成管状结构。肝素组的管状结构比对照组多 ,而肝素加bFGF组的管状结构比肝素组和bFGF组多。在Matrigel基膜基质凝胶上培养的内皮细胞向凝胶上层迁移 ,形成管状结构。管状结构较细 ,细胞较长。管状结构呈明显的牵拉状态。透射电镜下 ,淋巴管内皮细胞形成的管状结构具有毛细淋巴管的形态学特征。 结论 狗胸导管内皮细胞在Ⅰ型胶原蛋白凝胶和Matrigel基膜基质凝胶中形成毛细淋巴管样结构。细胞外基质对于淋巴管内皮细胞的形态变化、细胞迁移和淋巴管新生具有促进作用。
Objective To investigate the effect of extracellular matrix on lymphangiogenesis. METHODS: Dog thoracic duct endothelial cells were cultured on collagen type I collagen gel and Matrigel-based membrane matrix gel. The morphology of tubular structures formed by endothelial cells was observed under an inverted phase contrast microscope. The lymphatic-like structures were observed under transmission electron microscope. Instrument for the tubular structure for quantitative analysis. As a result, lymphatic endothelial cells grew into a monolayer on collagen type I collagen, but no tubular structure was observed. After the cells are covered with an upper gel, the endothelial cells undergo morphological changes to form a tubular structure. The tubular structure of heparin group than the control group, and heparin plus bFGF group tubular structure than the heparin group and bFGF group more. Endothelial cells cultured on Matrigel basement membrane gels migrate to the upper layer of the gel, forming a tubular structure. Tubular structure is thin, longer cells. Tubular structure was significantly pulled state. Under the transmission electron microscope, the tubular structure formed by lymphatic endothelial cells has morphological features of lymphatic vessels. Conclusion The dog thoracic ductal endothelial cells form lymphangioid-like structures in collagen type I collagen and Matrigel-based matrix hydrogels. Extracellular matrix can promote the morphological changes of lymphatic endothelial cells, cell migration and lymphangiogenesis.