title: Mimicking Metastases Including Tumor Stroma: A New Technique to Generate a Three-Dimensional Colorectal Cancer Model Based on a Biological Decellularized Intestinal Scaffold.
abstract: Tumor models based on cancer cell lines cultured two-dimensionally (2D) on plastic lack histological complexity and functionality compared to the native microenvironment. Xenogenic mouse tumor models display higher complexity but often do not predict human drug responses accurately due to species-specific differences. We present here a three-dimensional (3D) in vitro colon cancer model based on a biological scaffold derived from decellularized porcine jejunum (small intestine submucosa+mucosa, SISmuc). Two different cell lines were used in monoculture or in coculture with primary fibroblasts. After 14 days of culture, we demonstrated a close contact of human Caco2 colon cancer cells with the preserved basement membrane on an ultrastructural level as well as morphological characteristics of a well-differentiated epithelium. To generate a tissue-engineered tumor model, we chose human SW480 colon cancer cells, a reportedly malignant cell line. Malignant characteristics were confirmed in 2D cell culture: SW480 cells showed higher vimentin and lower E-cadherin expression than Caco2 cells. In contrast to Caco2, SW480 cells displayed cancerous characteristics such as delocalized E-cadherin and nuclear location of β-catenin in a subset of cells. One central drawback of 2D cultures-especially in consideration of drug testing-is their artificially high proliferation. In our 3D tissue-engineered tumor model, both cell lines showed decreased numbers of proliferating cells, thus correlating more precisely with observations of primary colon cancer in all stages (UICC I-IV). Moreover, vimentin decreased in SW480 colon cancer cells, indicating a mesenchymal to epithelial transition process, attributed to metastasis formation. Only SW480 cells cocultured with fibroblasts induced the formation of tumor-like aggregates surrounded by fibroblasts, whereas in Caco2 cocultures, a separate Caco2 cell layer was formed separated from the fibroblast compartment beneath. To foster tissue generation, a bioreactor was constructed for dynamic culture approaches. This induced a close tissue-like association of cultured tumor cells with fibroblasts reflecting tumor biopsies. Therapy with 5-fluorouracil (5-FU) was effective only in 3D coculture. In conclusion, our 3D tumor model reflects human tissue-related tumor characteristics, including lower tumor cell proliferation. It is now available for drug testing in metastatic context-especially for substances targeting tumor-stroma interactions.
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Sarah
authors.initials: S
authors.lastname: Nietzer
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Florentin
authors.initials: F
authors.lastname: Baur
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Stefan
authors.initials: S
authors.lastname: Sieber
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Jan
authors.initials: J
authors.lastname: Hansmann
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Thomas
authors.initials: T
authors.lastname: Schwarz
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Carolin
authors.initials: C
authors.lastname: Stoffer
authors.affiliation: 2 Translational Center Würzburg "Regenerative Therapies in Oncology and Musculoskeletal Disease, " Fraunhofer Institute Interfacial Engineering and Biotechnology IGB , Würzburg, Germany .
authors.forename: Heide
authors.initials: H
authors.lastname: Häfner
authors.affiliation: 3 Department of Surgery I, Molecular Oncology and Immunology, University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Martin
authors.initials: M
authors.lastname: Gasser
authors.affiliation: 3 Department of Surgery I, Molecular Oncology and Immunology, University Hospital of the Julius-Maximilians University , Würzburg, Germany .
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Waaga-Gasser
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .|2 Translational Center Würzburg "Regenerative Therapies in Oncology and Musculoskeletal Disease, " Fraunhofer Institute Interfacial Engineering and Biotechnology IGB , Würzburg, Germany .
authors.forename: Heike
authors.initials: H
authors.lastname: Walles
authors.affiliation: 1 Institute of Tissue Engineering and Regenerative Medicine (TERM), University Hospital of the Julius-Maximilians University , Würzburg, Germany .|2 Translational Center Würzburg "Regenerative Therapies in Oncology and Musculoskeletal Disease, " Fraunhofer Institute Interfacial Engineering and Biotechnology IGB , Würzburg, Germany .
authors.forename: Gudrun
authors.initials: G
authors.lastname: Dandekar
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D018920:Coculture Techniques; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000556:secondary*; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology*; D008954:Models, Biological; D012867:Skin / Q000378:metabolism / Q000473:pathology*; D017154:Stromal Cells / Q000378:metabolism / Q000473:pathology*; D013552:Swine; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D059016:Tumor Microenvironment