人胚肺成纤维细胞饲养兔角膜上皮和内皮细胞的体外培养

In vitra culture of rabbit corneal epithelium and endothelium supporting by human fetal lung fibroblast feeder cells

目的 研究人胚肺成纤维饲细胞体外培养兔角膜上皮和内皮细胞。方法 人胚肺成纤维细胞系细胞，经丝裂霉素Ｃ处理成饲细胞。兔角膜上皮和内皮细胞组织块原代培养，消化接种于人胚肺饲细胞瓶传代培养。体外培养的角膜上皮和内皮细胞行常规病理形态学检查，角膜上皮细胞行角蛋白、内皮细胞行神经烯醇化酶免疫组化染色检测。结果人胚肺成纤维细胞形态均一，易于分辨，经丝裂霉素处理后失去增殖能力。角膜上皮和内皮细胞原代培养５～７天，细胞密集，经人胚肺饲细胞共培养５代，细胞无衰老现象。检测角膜上皮细胞角蛋白、内皮细胞神经烯醇化酶表达阳性，人胚肺饲细胞表达阴性。结论 人胚肺饲细胞能够明显增强角膜上皮和内皮细胞的体外生存能力，提高细胞的增殖能力。

Objective To investigate properties of culture of rabbit corneal epithelium and endothelium in presence of human fetal lung fibroblast(hFLP) feeder cells in vitra. Methods Treated with Mitomycin C, hFLP became feeder cells. After primary culture,rabbit corneal epithelium and endothelium were digested and inoculated in bottle having hFLP feeder cells for serial passage. They were examined by routine pathology and anti-pan cytokeratin to epithelia and anti-neuron special enlose to endothelia immunohistochemistry. Results hFLP was resembling by itself in shape and could be simply distinguished in appearance. If treated by Mitomycin C, it lose proliferative capacity. After having primarily cultivated, the density of corneal epithelia and endothelia were thick and fast. When having co-cultivated with hFLP feeder cells for 5 passages, there was no caducity in corneal epithelia and endothelia. All of the third corneal epithelia expressed keratin and endothelia neuron special enlose, but hFLP did not express both. Conclusion hFLP feeder cells can enforce the survival and proliferative capacity of rabbit corneal epithelium and endothelium in vitra.