Kolostova Katarina, Zhang Yong, Hoffman Robert M, Bobek Vladimir
Department of Laboratory Genetics, University Hospital Kralovske Vinohrady, Srobarova 50, 100 34, Prague, Czech Republic.
J Fluoresc. 2014 Sep;24(5):1531-6. doi: 10.1007/s10895-014-1439-3. Epub 2014 Aug 22.
In the present study, we demonstrate an animal model and recently introduced size-based exclusion method for circulating tumor cells (CTCs) isolation. The methodology enables subsequent in vitro CTC-culture and characterization. Human lung cancer cell line H460, expressing red fluorescent protein (H460-RFP), was orthotopically implanted in nude mice. CTCs were isolated by a size-based filtration method and successfully cultured in vitro on the separating membrane (MetaCell®), analyzed by means of time-lapse imaging. The cultured CTCs were heterogeneous in size and morphology even though they originated from a single tumor. The outer CTC-membranes were blebbing in general. Abnormal mitosis resulting in three daughter cells was frequently observed. The expression of RFP ensured that the CTCs originated from lung tumor. These readily isolatable, identifiable and cultivable CTCs can be used to characterize individual patient cancers and for screening of more effective treatment.
在本研究中，我们展示了一种动物模型以及最近引入的基于大小的循环肿瘤细胞（CTC）分离排除方法。该方法能够进行后续的体外CTC培养和表征。将表达红色荧光蛋白的人肺癌细胞系H460（H460-RFP）原位植入裸鼠体内。通过基于大小的过滤方法分离CTC，并成功在分离膜（MetaCell®）上进行体外培养，通过延时成像进行分析。尽管培养的CTC源自单一肿瘤，但它们在大小和形态上是异质的。一般来说，CTC的外膜会出现气泡。经常观察到导致三个子细胞的异常有丝分裂。RFP的表达确保了CTC源自肺肿瘤。这些易于分离、鉴定和培养的CTC可用于表征个体患者的癌症，并用于筛选更有效的治疗方法。
