Abstract 1050: Efficient derivation and expansion of tumor cell lines from primary and xenotransplanted pancreatic tumors

Abstract

Cancer cell lines are used to study tumor biology and as models for testing of novel anti-cancer therapeutics. The vast majority of this work has been done using established cell lines that have been cultured for decades. Upon extensive in vitro propagation, cancer cell lines acquire multiple genetic and epigenetic alterations, lose initial heterogeneity present in the parental tumor, and tend to lack tumor initiating as well as multi-lineage differentiation capacity. Consequently, it has been shown that ‘classical' tumor cell lines only insufficiently resemble the characteristics of a tumor. In order to generate improved models for cancer research, novel cell lines are derived from primary tissues and used at low passages. However, this process is very inefficient for most tumor entities. In addition, most of the media used include largely undefined serum, such as FBS, which has been shown to drive primary tumor cell cultures to a more differentiated state when used over multiple passages. We have developed an advanced, serum-free medium for derivation and expansion of tumor cell lines from pancreatic tumors (Noll et al., 2016). This medium has been further optimized concerning formulation, stability, and usability. It allows for efficient generation of primary cell lines from both, patient and xenotransplanted tumors. Primary cell lines derived with this medium closely resembled essential characteristics of the parental tumor, including expression of subtype-specific markers, cellular heterogeneity, as well as genetic and epigenetic signatures. The derived cell lines have been used for genetic engineering and the developed medium allowed for the subsequent expansion of single-cell clones, a pre-requisite for reliable downstream assays. Importantly, culturing with our medium retained the tumorigenic potential as shown by xenotransplantation in immunodeficient mice. The resulting tumors closely and reliably resembled the initial patient tumor. This has been shown on the histomorphological as well as functional level, for example by conservation of tumor specific resistance mechanisms. Taken together, we have developed a serum-free medium for efficient derivation and expansion of tumor cell lines from primary and xenotransplanted pancreatic tumors, allowing for the establishment of easily accessible in vitro models. Moreover, the cell lines could also be used to generate corresponding xenograft models facilitating the translation of in vitro findings directly into a pre-clinical in vivo setting. Citation Format: David Agorku, Anne Langhammer, Elisa M. Noll, Christian Eisen, Silke Schult, Andreas Bosio, Martin R. Sprick, Andreas Trumpp, Olaf Hardt. Efficient derivation and expansion of tumor cell lines from primary and xenotransplanted pancreatic tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1050.