目的观察克隆形态是否与肿瘤细胞的不断自我更新相关。方法将A549细胞株单细胞悬液接种到96孔板,观察克隆形成率和克隆形态及传代情况,测量克隆大小,并计数细胞数目。分别观察早期、中间和晚期的黏附细胞数目,检测其细胞增殖活性和细胞免疫表型。结果A549细胞可形成3种类型的克隆集落,其中,Holoclone型占总克隆数的4％,克隆体积大,可连续传代20代以上,并可分化为其他2种类型的集落,增殖活性不随时间延长而降低,并表达干细胞标记。结论A549中Holoclone型集落存在不断自我更新的肿瘤干细胞。 Objective To observe whether the colony morphology is related to the continuous self-renewal of tumor cells. Methods The single cell suspension of A549 cells was inoculated into 96-well plates. The formation rate of clones, the morphology and passage of clones were observed, the size of clones was measured, and the number of cells was counted. The numbers of early, middle and late adherent cells were observed, and their cell proliferative activity and cellular immunophenotype were detected. Results Three types of colonies were formed in A549 cells. Among them, Holoclone accounted for 4% of the total number of clones. The clone volume was large and could be continuously passaged for more than 20 passages and differentiated into two other types of colonies. The proliferative activity did not change with time Lengthened and decreased, and expressed stem cell markers. Conclusion The Holoclone-type colonies in A549 have continuously self-renewing tumor stem cells.