title: Bioreactor-engineered cancer tissue-like structures mimic phenotypes, gene expression profiles and drug resistance patterns observed "in vivo".
abstract: Anticancer compound screening on 2D cell cultures poorly predicts "in vivo" performance, while conventional 3D culture systems are usually characterized by limited cell proliferation, failing to produce tissue-like-structures (TLS) suitable for drug testing. We addressed engineering of TLS by culturing cancer cells in porous scaffolds under perfusion flow. Colorectal cancer (CRC) HT-29 cells were cultured in 2D, on collagen sponges in static conditions or in perfused bioreactors, or injected subcutaneously in immunodeficient mice. Perfused 3D (p3D) cultures resulted in significantly higher (p < 0.0001) cell proliferation than static 3D (s3D) cultures and yielded more homogeneous TLS, with morphology and phenotypes similar to xenografts. Transcriptome analysis revealed a high correlation between xenografts and p3D cultures, particularly for gene clusters regulating apoptotic processes and response to hypoxia. Treatment with 5-Fluorouracil (5-FU), a frequently used but often clinically ineffective chemotherapy drug, induced apoptosis, down-regulation of anti-apoptotic genes (BCL-2, TRAF1, and c-FLIP) and decreased cell numbers in 2D, but only "nucleolar stress" in p3D and xenografts. Conversely, BCL-2 inhibitor ABT-199 induced cytotoxic effects in p3D but not in 2D cultures. Our findings advocate the importance of perfusion flow in 3D cultures of tumor cells to efficiently mimic functional features observed "in vivo" and to test anticancer compounds.
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Hirt
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Adam
authors.initials: A
authors.lastname: Papadimitropoulos
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Manuele G
authors.initials: MG
authors.lastname: Muraro
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Valentina
authors.initials: V
authors.lastname: Mele
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Evangelos
authors.initials: E
authors.lastname: Panopoulos
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Cremonesi
authors.affiliation: Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Robert
authors.initials: R
authors.lastname: Ivanek
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Elke
authors.initials: E
authors.lastname: Schultz-Thater
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland.
authors.forename: Raoul A
authors.initials: RA
authors.lastname: Droeser
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Chantal
authors.initials: C
authors.lastname: Mengus
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Michael
authors.initials: M
authors.lastname: Heberer
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland.
authors.forename: Daniel
authors.initials: D
authors.lastname: Oertli
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Giandomenica
authors.initials: G
authors.lastname: Iezzi
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland.
authors.forename: Paul
authors.initials: P
authors.lastname: Zajac
authors.affiliation: Institute of Pathology, University of Basel, Switzerland.
authors.forename: Serenella
authors.initials: S
authors.lastname: Eppenberger-Castori
authors.affiliation: Institute of Pathology, University of Basel, Switzerland.
authors.forename: Luigi
authors.initials: L
authors.lastname: Tornillo
authors.affiliation: Institute of Pathology, University of Basel, Switzerland.
authors.forename: Luigi
authors.initials: L
authors.lastname: Terracciano
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland. Electronic address: ivan.martin@usb.ch.
authors.forename: Ivan
authors.initials: I
authors.lastname: Martin
authors.affiliation: Department of Surgery, University Hospital Basel, Switzerland; Department of Biomedicine, University of Basel, Switzerland. Electronic address: giulio.spagnoli@usb.ch.
authors.forename: Giulio C
authors.initials: GC
authors.lastname: Spagnoli
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000627:therapeutic use; D061249:Batch Cell Culture Techniques / Q000295:instrumentation; D032701:Biomimetics / Q000379:methods; D019149:Bioreactors; D049109:Cell Proliferation / Q000187:drug effects; D019008:Drug Resistance, Neoplasm; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005472:Fluorouracil / Q000627:therapeutic use*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D019073:HT29 Cells; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism; D009374:Neoplasms, Experimental / Q000188:drug therapy / Q000473:pathology* / Q000503:physiopathology*; D010641:Phenotype; D023822:Tissue Engineering / Q000295:instrumentation*
chemical_list: D000964:Antimetabolites, Antineoplastic; D009363:Neoplasm Proteins; D005472:Fluorouracil
keywords: Bioreactors; Colorectal cancer; Drug resistance; Tri-dimensional cultures; Tumor tissue-like structures