Abstract 4258: Multi-tumor cell culture medium supports a high take rate and improves culture growth rate in five tumor types
Authors: Agoston Elin S. , Novak Marian , Salimi Naghmeh , Chao Alex , Kent Jeffrey , Agoston Agoston T. , Drapkin Ronny 
Published: July 2016

Abstract
The efficiency of primary cell line derivation is inadequate, which presents a substantial hurdle in the study of many solid tumor types. We examined whether substitution of a novel multi-tumor cell culture medium into existing cell culture protocols may improve the efficiency of primary cell culture and cell line derivation.
The multi-tumor medium RETM (Renaissance Essential Tumor Medium) was initially screened for the ability to achieve primary, extended, and continuous cultures from solid tumors of the breast, lung, colon, prostate and ovary (serous and endometrioid). A pan-cytokeratin antibody was then used to confirm epithelial origin of the expanded cell lines. Performance of RETM was compared to two established media types (RPMI, and Fallopian Tube Medium) in extended primary culture of five high-grade serous carcinoma (HGSC) tumor samples isolated from patients’ ascites. Immunofluorescent staining of Pax8, an important Mullerian lineage marker, was used to confirm the purity and the expected lineage identity of the resulting cell lines. To examine genetic drift, we performed SNP genotyping on the original tumor and the cultured cells in our extended primary cultures of HGSC and our continuous cultures from breast, lung, and colon. The resulting genotype was then compared to the original tumor and the percent matching genotype was determined.
Of the six tumor types screened, five grew as long term cultures. Without additional optimization of methodology in HGSC, RETM supported a higher take rate (extended culture in 4 of 5 isolates versus 1 of 5 for established media types) as well as a 10-fold higher average growth rate in the same four cases (RETM doublings/day: 0.309 versus 0.030 for RPMI [p = 0.004] and 0.023 for FTM [p = 0.001]). In all cases, fibroblast overgrowth was ruled out by positive cytokeratin staining. The genetic analysis demonstrated a high degree of fidelity in genotypes between the patients’ tumors and the corresponding cell lines as follows: DF30 (HGSC at 23 doublings) 99.7%; DF68 (HGSC at 25 doublings) 95.5%; Wood (breast ductal and lobular carcinoma at 150 doublings) 98.3%; Jacket (lung adenocarcinoma at 150 doublings) 87.2%; and Ferry (colon adenocarcinoma at 30 doublings) 99.6%.
We have shown for the first time that a single cell culture medium can support establishment of long-term cell lines established from a number of different tumor types including breast, lung, colon, endometrioid ovarian, and HGSC. Our results demonstrate that, compared to the current standard, RETM significantly improves both culture take rate and growth rate of the extended HGSC primary cultures. Together with the absence of cell culture crisis, the maintenance of genotype across a large number of population doublings indicate these cell lines are genetically stable and may be cultured to higher passages with reduced concern for genetic drift.
Citation Format: Elin S. Agoston, Marian Novak, Naghmeh Salimi, Alex Chao, Jeffrey Kent, Agoston T. Agoston, Ronny Drapkin. Multi-tumor cell culture medium supports a high take rate and improves culture growth rate in five tumor types. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4258.