title: Human Microtumors Generated in 3D: Novel Tools for Integrated In Situ Studies of Cancer Immunotherapies.
abstract: Cellular immunotherapy targeting human tumor antigens is a promising strategy to treat solid tumors. Yet clinical results of cellular immunotherapy are disappointing. Moreover, the currently available in vitro human tumor models are not designed to study the optimization of T-cell therapies of solid tumors. Here, we describe a novel assay for multiparametric in situ analysis of therapeutic effects on individual human three-dimensional (3D) tumors. In this assay, tumors of several millimeter diameter are generated from human cancer cell lines of different tumor entities in a collagen type I microenvironment. A newly developed approach for efficient morphological analysis reveals that these in vitro tumors resemble many characteristics of the corresponding clinical cancers such as histological features, immunohistochemical staining patterns, distinct tumor growth compartments and heterogeneous protein expression. To assess the response to therapy with tumor antigen specific T-cells, standardized protocols are described to determine T-cell infiltration and tumor destruction by monitoring soluble factors and tumor growth. Human tumors engineered in 3D collagen scaffolds are excellent in vitro surrogates for avascular tumor stages allowing integrated analyses of the antitumor efficacy of cancer specific immunotherapy in situ.
authors.affiliation: Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leident, The Netherlands. hambach.lothar@mh-hannover.de.|Department of Hematology, Hemostasis, Oncology and Stem cell transplantation Hannover Medical School, Carl-Neuberg-Str. 1, Hannover, 30625, Germany. hambach.lothar@mh-hannover.de.
authors.forename: Lothar
authors.initials: L
authors.lastname: Hambach
authors.affiliation: Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leident, The Netherlands.|Blood Transfusion Centre, Swiss Red Cross, Basel, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Buser
authors.affiliation: Department of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Vermeij
authors.affiliation: Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leident, The Netherlands.
authors.forename: Nadine
authors.initials: N
authors.lastname: Pouw
authors.affiliation: Department of Pathology, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Theo
authors.initials: T
authors.lastname: van der Kwast
authors.affiliation: Department of Immunohaematology and Blood Transfusion, Leiden University Medical Center, Leident, The Netherlands.
authors.forename: Els
authors.initials: E
authors.lastname: Goulmy
mesh_terms: D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy; D009369:Neoplasms / Q000628:therapy*; D016612:Paraffin Embedding; D018874:Spheroids, Cellular / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism; D023822:Tissue Engineering
chemical_list: D016207:Cytokines
keywords: Cell adhesion and extracellular matrix; In vitro tumor model; Multiparametric assay; Novel assay technology; Targeted therapies