title: A computer-assisted 3D model for analyzing the aggregation of tumorigenic cells reveals specialized behaviors and unique cell types that facilitate aggregate coalescence.
abstract: We have developed a 4D computer-assisted reconstruction and motion analysis system, J3D-DIAS 4.1, and applied it to the reconstruction and motion analysis of tumorigenic cells in a 3D matrix. The system is unique in that it is fast, high-resolution, acquires optical sections using DIC microscopy (hence there is no associated photoxicity), and is capable of long-term 4D reconstruction. Specifically, a z-series at 5 μm increments can be acquired in less than a minute on tissue samples embedded in a 1.5 mm thick 3D Matrigel matrix. Reconstruction can be repeated at intervals as short as every minute and continued for 30 days or longer. Images are converted to mathematical representations from which quantitative parameters can be derived. Application of this system to cancer cells from established lines and fresh tumor tissue has revealed unique behaviors and cell types not present in non-tumorigenic lines. We report here that cells from tumorigenic lines and tumors undergo rapid coalescence in 3D, mediated by specific cell types that we have named "facilitators" and "probes." A third cell type, the "dervish", is capable of rapid movement through the gel and does not adhere to it. These cell types have never before been described. Our data suggest that tumorigenesis in vitro is a developmental process involving coalescence facilitated by specialized cells that culminates in large hollow spheres with complex architecture. The unique effects of select monoclonal antibodies on these processes demonstrate the usefulness of the model for analyzing the mechanisms of anti-cancer drugs.
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Amanda
authors: A
authors: Scherer
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Spencer
authors: S
authors: Kuhl
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Deborah
authors: D
authors: Wessels
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Daniel F
authors: DF
authors: Lusche
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Brett
authors: B
authors: Hanson
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Joseph
authors: J
authors: Ambrose
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: Edward
authors: E
authors: Voss
authors: Mercy Hospital System of Des Moines, Des Moines, Iowa, United States of America.
authors: Emily
authors: E
authors: Fletcher
authors: Mercy Hospital System of Des Moines, Des Moines, Iowa, United States of America.
authors: Charles
authors: C
authors: Goldman
authors: Monoclonal Antibody Research Institute, Developmental Studies Hybridoma Bank, Department of Biology, University of Iowa, Iowa City, Iowa, 52242, United States of America.
authors: David R
authors: DR
authors: Soll
mesh_terms: D002449:Cell Aggregation / Q000502:physiology*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D003094:Collagen; D004338:Drug Combinations; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007797:Laminin; D008954:Models, Biological; D009369:Neoplasms / Q000503:physiopathology*; D011509:Proteoglycans; D059008:Time-Lapse Imaging / Q000379:methods*
chemical_list: D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen