Ex vivo organotypic culture system of precision-cut slices of human pancreatic ductal adenocarcinoma

Abstract

Pancreatic ductal adenocarcinoma (PDAC) has a poor prognosis, which is mainly due to late diagnosis and profound resistance to treatment. The latter is to a large extent attributed to the tumor stroma that is exceedingly prominent in PDAC and engages in complex interactions with the cancer cells. Hence, relevant preclinical models of PDAC should also include the tumor stroma. We herein describe the establishment and functional validation of an ex vivo organotypic culture of human PDAC that is based on precision-cut tissue slices from surgical specimens and reproducibly recapitulates the complex cellular and acellular composition of PDAC, including its microenvironment. The cancer cells, tumor microenvironment and interspersed remnants of nonneoplastic pancreas contained in these 350 µm thick slices maintained their structural integrity, phenotypic characteristics and functional activity when in culture for at least 4 days. In particular, tumor cell proliferation persisted and the grade of differentiation and morphological phenotype remained unaltered. Cultured tissue slices were metabolically active and responsive to rapamycin, an mTOR inhibitor. This culture system is to date the closest surrogate to the parent carcinoma and harbors great potential as a drug sensitivity testing system for the personalized treatment of PDAC.

Optimization of Organotypic Cultures of Mouse Spleen for Staining and Functional Assays

“…Organotypic culture has been extended to several other organs of the neuroendocrine system (1) and, more recently, to tumor-derived tissues (3). Of note, only one report describes the application of this technique to lymphoid tissues of human origin (4), notwithstanding the wealth of information generated over the last decade on the complex interactions that occur among immune, stromal, and cancer cells (5,6).…”

Ex-Vivo Treatment of Tumor Tissue Slices as a Predictive Preclinical Method to Evaluate Targeted Therapies for Patients with Renal Carcinoma

“…Given the variety of mechanisms triggered by molecularly targeted agents in cancers and their late-stage clinical trials, the validation of drug sensitivity predictive models may be critical to identify the right drug for the right patients and help to understand determinants of responsiveness, wherein alternative treatments could potentially overcome resistance [45]. There is a recently growing body of literature describing PDTSC from different normal and tumor tissues [29][30][31][32][33][34][35][36][37][38]. However, to our knowledge, the present study is the first to demonstrate the potential use of this approach to evaluate renal cancer response to novel therapies while modeling the tumor immune microenvironment.…”

“…We set out to determine whether an ex-vivo treatment protocol could be used as a means of determining ccRCC sensitivity to various cytotoxic agents. The PDTSC methodology has been previously used to evaluate the drug sensitivity of normal and tumor tissues [29][30][31][32][33][34][35][36][37][38]. Therefore, we set up an adaptation of this method outlined in Figure 2A, as an ex-vivo protocol to examine responses of ccRCC to different therapeutic agents.…”

“…To address these challenges, ex-vivo culture of intact tumor slices is potentially an extremely attractive system that has been already validated in various types of cancers [29][30][31][32][33][34][35][36][37][38].…”

Optimal, Large-Scale Propagation of Mouse Mammary Tumor Organoids

“…Tumor organoids can be isolated from experimental mouse models as well as human patient primary and metastatic tumors, allowing for the representation of a wide range of tumor phenotypes and genotypes. To observe and perturb complex interactions amongst tumor cells, normal cells, and stroma, multicellular tumor organoids are embedded in a variety of 3D matrix platforms [2,11,12,20,32,[43][44][45]. In this way, investigators have used tumor organoids in 3D culture to uncover essential aspects of tumor formation, met a s t a s i s , i n t e r c e l l u l a r t u m o r -t u m o r a n d t u m o rmicroenvironmental interactions, and therapeutic drug resistance [4-6, 10, 24, 46, 61].…”