Effects of extensive passaging on the characteristics of pancreatic ductal adenocarcinoma cancer cell line HG008-T

Abstract Pancreatic cancer is one of the major causes of cancer death. Pancreatic cancer cell lines are important for drug and cancer treatment development. In a collaborative effort, we established a novel Cancer Genome-in-a-Bottle cell line HG008-T from a pancreatic cancer patient explicitly consented to make public extensive genomic data, which was recently published. The HG008-T cell line is publicly available at ATCC as CRL-3734. Characterizing a cell line before it is available from a public repository is critical to ensure scientific reliability, and reproducibility. The cell line has known somatic variants in KRAS , TP53 , SMAD4 , and CDKN2A genes, and was derived from a patient who received platinum neoadjuvant therapy. To complement the extensive genomic characterization published recently, in this study we characterized HG008-T over 80 passages by comparing early-passage cells with late-passage cells. The cells were found to proliferate faster after extensive passaging and lost heterogeneity in morphology. Following extensive passaging, an increased proportion of cells exhibited whole-genome doubling (WGD), which may reflect the selective expansion of WGD clones due to enhanced proliferative capacity and/or de novo genome doubling events occurring in non-WGD cells during prolonged culture. The late-passage cells exhibited increased anchorage-independent growth. No significant surface marker expression alteration was observed. However, following approximately 80 passages, heterogeneous alterations in marker gene expression were observed. The late-passage cells were more sensitive to cisplatin but exhibited a modest shift in sensitivity to the CDK4/6 inhibitor palbociclib. In conclusion, extensive passaging altered certain characteristics of HG008-T cells, therefore, passage history should be indicated in future studies using HG008-T cell line as a cancer cell model.