利用气液界面无血清培养原代兔气管上皮细胞的研究

STUDIES ON PRIMARY CULTURE OF RABBIT TRACHEAL EPITHELIAL CELLS IN SERUM-FREE HORMONESUPPLEMENTED MEDIUM IN A BIPHASIC CHAMBER SYSTEM

用低温酶消化法分离兔气管上皮细胞,具有细胞损伤小,活力及纯度高的优点,成纤维细胞污染极低。人胎盘胶原提高了气管上皮细胞贴壁性。无血清培养基能促进细胞增殖,分化和成熟。气液界面培养方式更好地模拟了气管上皮细胞的天然生长环境,在膜上呈复层生长,有利于细胞的分化成熟及功能表达。光镜下细胞形态及免疫组化细胞角蛋白染色阳性证实培养细胞为气管上皮细胞。本文所建立的兔气管上皮细胞体外气液界面无血清培养方法为研究气 管上皮细胞的生理和病理提供了一个十分有用的模型。

In ordcr to study the physiology and pathophysiology of tracheal epithelium.a primary culture of tracheal epithelial cells at air-liquid interface has been established in our laboratory. Rabbit tracheal epithelia were isolated by using low temperature protease digestion, which was not harmful to the cell viability and could almost eliminate fibrob-lastlike cells. Coating human placenta collagen was beneficial to the attachrnent of epithelial celis. Serum-free hor-mone-supplemented medium could promote cell multiplication and differentiation. Biphasic chamber system provided an air-liquid interface, an environment closely resembling to the in vivo situation,allowing multicellular layers of ep-ithelium to grow. The nature of the tracheal epithelial cells was confirmed by morphology and immunocytochemistry. This culture system provided an ideal model for more sophisticated in vitro modelling of in vivo functions and thus would be useful in the study of physiology and pathophysiology of tracheal epithelium.