Rafferty K A, Ruben R L, Young S K
In Vitro. 1978 Feb;14(2):227-35. doi: 10.1007/BF02618227.
Normal cultures of epithelial appearance were initiated by trypsinization of a surgically resected, histologically normal branchial cyst. Cellular morphology was consistent with derivation from the stratified squamous epithelium of the cyst or from vascular endothelium, although electron micrographs of the cultured cells failed to show any junctional complexes. Infection with SV40 produced transformants which were also epithelioid in appearance. These grew vigorously for 22 to 50 population doublings (about 23 to 32 subcultures, depending upon regimen) and then became quiescent. During this evolution, virus was detectable at all stages by both direct isolation (cell extracts) and cocultivation with permissive cells. In two sublines in which selection for rapidly growing cell types occureed, virus was detected only by cocultivation. The work confirms that of others in the finding that normal human epithelial cells are susceptible to transformation by oncogenic viruses, but are apparently less responsive than are fibroblasts to such transforming agents. It also suggests that subcultivation techniques that maintain the populations of transformed cells at low density tend to select against cell strains that are continous producers of infectious virus.
通过对手术切除的组织学正常的鳃裂囊肿进行胰蛋白酶消化,启动了具有上皮外观的正常培养。细胞形态与囊肿的复层鳞状上皮或血管内皮来源一致,尽管培养细胞的电子显微镜照片未显示任何连接复合体。用SV40感染产生的转化体外观也是上皮样的。这些转化体旺盛生长22至50代(约23至32次传代,取决于培养方案),然后进入静止期。在这个过程中,通过直接分离(细胞提取物)和与允许细胞共培养,在所有阶段都可检测到病毒。在两个进行快速生长细胞类型选择的亚系中,仅通过共培养检测到病毒。这项工作证实了其他人的发现,即正常人类上皮细胞易被致癌病毒转化,但显然比成纤维细胞对这种转化剂的反应性低。它还表明,将转化细胞群体维持在低密度的传代培养技术倾向于选择不连续产生感染性病毒的细胞株。