title: Temperature responsive hydrogels enable transient three-dimensional tumor cultures via rapid cell recovery.
abstract: Recovery of live cells from three-dimensional (3D) culture would improve analysis of cell behaviors in tissue engineered microenvironments. In this work, we developed a temperature responsive hydrogel to enable transient 3D culture of human glioblastoma (GBM) cells. N-isopropylacrylamide was copolymerized with hydrophilic grafts and functionalized with the cell adhesion peptide RGD to yield the novel copolymer poly(N-isopropylacrylamide-co-Jeffamine(®) M-1000 acrylamide-co-hydroxyethylmethacrylate-RGD), or PNJ-RGD. This copolymer reversibly gels in aqueous solutions when heated under normal cell culture conditions (37°C). Moreover, these gels redissolve within 70 s when cooled to room temperature without the addition of any agents to degrade the synthetic scaffold, thereby enabling rapid recollection of viable cells after 3D culture. We tested the efficiency of cell recovery following extended 3D culture and were able to recover more than 50% of viable GBM cells after up to 7 days in culture. These data demonstrate the utility of physically crosslinked PNJ-RGD hydrogels as a platform for culture and recollection of cells in 3D.
authors.affiliation: Barrow Brain Tumor Research Center, Barrow Neurological Institute, 350 W. Thomas Road, Neuroscience Research Center 441, Phoenix, Arizona, 85013.|School of Biological and Health Systems Engineering, Arizona State University, PO Box 879709, Tempe, Arizona, 85287.
authors.forename: John M
authors.identifier: http://orcid.org/0000-0002-0817-0966
authors.initials: JM
authors.lastname: Heffernan
authors.affiliation: Barrow Brain Tumor Research Center, Barrow Neurological Institute, 350 W. Thomas Road, Neuroscience Research Center 441, Phoenix, Arizona, 85013.
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Overstreet
authors.affiliation: Barrow Brain Tumor Research Center, Barrow Neurological Institute, 350 W. Thomas Road, Neuroscience Research Center 441, Phoenix, Arizona, 85013.|School of Biological and Health Systems Engineering, Arizona State University, PO Box 879709, Tempe, Arizona, 85287.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Srinivasan
authors.affiliation: School of Biological and Health Systems Engineering, Arizona State University, PO Box 879709, Tempe, Arizona, 85287.
authors.forename: Long D
authors.initials: LD
authors.lastname: Le
authors.affiliation: School of Biological and Health Systems Engineering, Arizona State University, PO Box 879709, Tempe, Arizona, 85287.
authors.forename: Brent L
authors.initials: BL
authors.lastname: Vernon
authors.affiliation: Barrow Brain Tumor Research Center, Barrow Neurological Institute, 350 W. Thomas Road, Neuroscience Research Center 441, Phoenix, Arizona, 85013.|School of Biological and Health Systems Engineering, Arizona State University, PO Box 879709, Tempe, Arizona, 85287.
authors.forename: Rachael W
authors.initials: RW
authors.lastname: Sirianni
mesh_terms: D000178:Acrylamides / Q000138:chemical synthesis / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002850:Chromatography, Gel; D004548:Elasticity; D006801:Humans; D020100:Hydrogels / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D012212:Rheology; D013696:Temperature; D054457:Tissue Scaffolds / Q000737:chemistry; D014407:Tumor Cells, Cultured; D014783:Viscosity
chemical_list: D000178:Acrylamides; D020100:Hydrogels; C067295:N-isopropylacrylamide
keywords: NIPAAm; RGD; cell recollection; glioblastoma; thermoreversible scaffold