Goldman W E, Baseman J B
In Vitro. 1980 Apr;16(4):313-9. doi: 10.1007/BF02618337.
A reliable cell isolation technique was developed to allow the cultivation of cells from the hamster respiratory tract. Repeated thermolysin treatments and gradient centrifugation yielded a cellculture completely free from contamination by fibroblasts. Viable cells could be isolated from as little tissue as a single hamster trachea, but in vitro proliferation occurred only if the hamster was less than 4 months of age. The cultured cells could be repeatedly passaged and subcultured for weeks by employing normal tissue culture techniques. Morphologically, the monolayers appeared to be a homogeneous population of epithelial cells, and successful cloning of freshly isolated single cells resulted in apparently identical cultures. The epiethelial origin of these cells was also suggested by continued growth in minimum essential medium with D-valine substituted for L-valine. The relative ease with which this cell type can be isolated, cultured, and manipulated in vitro should encourage its application as a model of the respiratory epithelium.
开发了一种可靠的细胞分离技术，以实现仓鼠呼吸道细胞的培养。反复进行胰蛋白酶处理和梯度离心，得到了完全没有成纤维细胞污染的细胞培养物。从少至单个仓鼠气管的组织中即可分离出活细胞，但只有当仓鼠年龄小于4个月时，体外增殖才会发生。通过采用常规组织培养技术，培养的细胞可以反复传代和继代培养数周。从形态学上看，单层细胞似乎是上皮细胞的同质群体，对新分离的单个细胞进行成功克隆可得到明显相同的培养物。用D-缬氨酸替代L-缬氨酸的最低必需培养基中细胞的持续生长也表明了这些细胞的上皮起源。这种细胞类型在体外相对容易分离、培养和操作，这应会促使其作为呼吸道上皮模型得到应用。
