Diglio C A, Kikkawa Y
Lab Invest. 1977 Dec;37(6):622-31.
Type II lung epithelial cells were isolated from rabbit lungs using the method of Kikkawa and Yoneda (Kikkawa Y, Yoneda K: Lab Invest 30: 76, 1974). Successful primary cultures were obtained only after utilizing high density cell plating (greater than 3 X 10(5) viable cells per sq. cm.) and allowing an attachment time of 48 hours. Attachment efficiency of the isolated cell preparations was highest when medium was supplemented with 10% fetal calf serum. These conditions enabled us to obtain consistently successful primary type II lung cell cultures. Cultures were monitored for a period of 2 weeks after initiation. Light, phase, and electron microscopy examination demonstrated that these primary cultures were indeed type II cells. The principal morphologic feature was the presence of dense lamellar granules in these cells. These primary cultures retained the characteristic type II features for 3 to 5 days in vitro, after which cultures exhibited a progressive deterioration and loss of their phenotypic properties. This behavioral pattern of type II cells in culture may represent both accelerated proliferation and accelerated transformation of these cells into type I epithelial cells.
采用Kikkawa和Yoneda的方法（Kikkawa Y, Yoneda K: Lab Invest 30: 76, 1974）从兔肺中分离出II型肺上皮细胞。仅在采用高密度细胞接种（每平方厘米大于3×10⁵个活细胞）并给予48小时的贴壁时间后，才成功获得原代培养物。当培养基中添加10%胎牛血清时，分离的细胞制剂的贴壁效率最高。这些条件使我们能够始终如一地成功获得II型肺细胞原代培养物。培养开始后监测2周。光学显微镜、相差显微镜和电子显微镜检查表明，这些原代培养物确实是II型细胞。主要形态学特征是这些细胞中存在致密板层小体。这些原代培养物在体外保留II型细胞特征3至5天，之后培养物表现出逐渐退化并丧失其表型特性。培养的II型细胞的这种行为模式可能代表这些细胞的加速增殖以及加速向I型上皮细胞的转化。
