title: A three dimensional micropatterned tumor model for breast cancer cell migration studies.
abstract: Breast cancer cell invasion is a highly orchestrated process driven by a myriad of complex microenvironmental stimuli, making it difficult to isolate and assess the effects of biochemical or biophysical cues (i.e. tumor architecture, matrix stiffness) on disease progression. In this regard, physiologically relevant tumor models are becoming instrumental to perform studies of cancer cell invasion within well-controlled conditions. Herein, we explored the use of photocrosslinkable hydrogels and a novel, two-step photolithography technique to microengineer a 3D breast tumor model. The microfabrication process enabled precise localization of cell-encapsulated circular constructs adjacent to a low stiffness matrix. To validate the model, breast cancer cell lines (MDA-MB-231, MCF7) and non-tumorigenic mammary epithelial cells (MCF10A) were embedded separately within the tumor model, all of which maintained high viability throughout the experiments. MDA-MB-231 cells exhibited extensive migratory behavior and invaded the surrounding matrix, whereas MCF7 or MCF10A cells formed clusters that stayed confined within the circular tumor regions. Additionally, real-time cell tracking indicated that the speed and persistence of MDA-MB-231 cells were substantially higher within the surrounding matrix compared to the circular constructs. Z-stack imaging of F-actin/α-tubulin cytoskeletal organization revealed unique 3D protrusions in MDA-MB-231 cells and an abundance of 3D clusters formed by MCF7 and MCF10A cells. Our results indicate that gelatin methacrylate (GelMA) hydrogel, integrated with the two-step photolithography technique, has great promise in the development of 3D tumor models with well-defined architecture and tunable stiffness.
authors.affiliation: School of Biological and Health Systems Engineering (SBHSE), Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Nitish
authors.initials: N
authors.lastname: Peela
authors.affiliation: School of Biological and Health Systems Engineering (SBHSE), Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Feba S
authors.initials: FS
authors.lastname: Sam
authors.affiliation: Center for Biological Physics, Arizona State University, Tempe, AZ 85287, USA; Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Wayne
authors.initials: W
authors.lastname: Christenson
authors.affiliation: School of Biological and Health Systems Engineering (SBHSE), Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Danh
authors.initials: D
authors.lastname: Truong
authors.affiliation: University of Arizona Cancer Center, Department of Cellular and Molecular Medicine, Tucson, AZ 85724, USA.
authors.forename: Adam W
authors.initials: AW
authors.lastname: Watson
authors.affiliation: University of Arizona Cancer Center, Department of Cellular and Molecular Medicine, Tucson, AZ 85724, USA.
authors.forename: Ghassan
authors.initials: G
authors.lastname: Mouneimne
authors.affiliation: Center for Biological Physics, Arizona State University, Tempe, AZ 85287, USA; Department of Physics, Arizona State University, Tempe, AZ 85287, USA; Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Ros
authors.affiliation: School of Biological and Health Systems Engineering (SBHSE), Arizona State University, Tempe, AZ 85287, USA. Electronic address: mnikkhah@asu.edu.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Nikkhah
mesh_terms: D000199:Actins / Q000378:metabolism; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D048430:Cell Shape; D002470:Cell Survival; D058948:Cell Tracking; D003599:Cytoskeleton / Q000378:metabolism; D005260:Female; D005453:Fluorescence; D006801:Humans; D007211:Indoles / Q000378:metabolism; D055616:Microtechnology / Q000379:methods*; D008954:Models, Biological; D012235:Rhodamines / Q000378:metabolism; D059008:Time-Lapse Imaging
chemical_list: D000199:Actins; D007211:Indoles; D012235:Rhodamines; C007293:DAPI; C029773:rhodamine B
keywords: Gelatin methacrylate (GelMA); MCF10A; MCF7; MDA-MB-231; Microengineering; Photolithography; Real-time cell tracking; Tumor model