Abstract
We have developed 3D-tumoroids and tumor slice in vitro culture systems from surgical tumor specimens derived from patients with colorectal cancer (CRC) or lung cancer to evaluate immune cell populations infiltrating cultured tissues. The system incorporates patient's peripherally and tumor-derived immune cells into tumoroid in vitro cultures to evaluate the ability of the culture to mimic an immunosuppressive tumor microenvironment (ITM). This system enables analysis of tumor response to standard therapy within weeks of surgical resection. Here we show that tumoroid cultures from a CRC patient are highly sensitive to the thymidylate synthase inhibitor 5-fluorouracil (adrucil) but less sensitive to the combination of nucleoside analog trifluridine and thymidine phosphorylase inhibitor tipiracil (Lonsurf). Moreover, re-introduction of isolated immune cells derived from surrounding and infiltrating tumor tissue as well as CD45+ tumor infiltrating hematopoietic cells displayed prolonged (>10 days) survival in co-culture. Established tumor slice cultures were found to contain both an outer epithelial and inner stromal cell compartment mimicking tumor structure in vivo.Collectively, these data suggest that, 3D-tumoroid and slice culture assays may provide a feasible in vitro approach to assess efficacy of novel therapeutics in the context of heterogeneous tumor-associated cell types including immune and non-transformed stromal cells. In addition, delineating the impact of therapeutics on immune cells, and cell types involved in therapeutic resistance mechanisms may be possible in general or for patient-specific responses.
Highlights
The successful establishment of exponentially growing viable cells from patient-derived tumors in in vitro cell cultures has been found to occur from a minority of tumor tissues following artificial selection of a sub-population of tumor cells
Recent advances in patient-derived 3D cultures have facilitated what is perhaps more relevant modeling of cancer in vitro and this can be combined with whole-genome sequencing approaches to couple functional and correlative tumorspecific data with the promise to shape the landscape of precision medicine. 3D in vitro tumoroid cultures can be rapidly established from biopsied or resected tumor tissues and subsequently subjected to in vitro sub-cultivation, expansion and genomic manipulation using e.g. CRISPR/ Cas9 methodology [2]
Upon troubleshooting the case (ReCa091516) from which we failed to prepare tumoroid and air-liquid interface (ALI) cultures it was discovered that the specimen that was processed only contained fibrous stroma and was completely devoid of tumor epithelium
Summary
The successful establishment of exponentially growing viable cells from patient-derived tumors in in vitro cell cultures has been found to occur from a minority of tumor tissues following artificial selection of a sub-population of tumor cells. Such limitations make it difficult to model patient variability in www.impactjournals.com/oncotarget drug responses in vitro. The methodology to establish tissue in vitro cultures has been available for more than a century [1], the use of two dimensional (2D) culture techniques to establish continuous tumor cell lines from explanted tumor tissue cultures is hampered by artificial culture conditions that severely diminish the success rate, and tumor clonal heterogeneity.
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