title: A novel 2.5D culture platform to investigate the role of stiffness gradients on adhesion-independent cell migration.
abstract: Current studies investigating the role of biophysical cues on cell migration focus on the use of culture platforms with static material parameters. However, migrating cells in vivo often encounter spatial variations in extracellular matrix stiffness. To better understand the effects of stiffness gradients on cell migration, we developed a 2.5D cell culture platform where cells are sandwiched between stiff tissue culture plastic and soft alginate hydrogel. Under these conditions, we observed migration of cells from the underlying stiff substrate into the alginate matrix. Observation of migration into alginate in the presence of integrin inhibition as well as qualitative microscopic analyses suggested an adhesion-independent cell migration mode. Observed migration was dependent on alginate matrix stiffness and the RhoA-ROCK-myosin-II pathway; inhibitors specifically targeting ROCK and myosin-II arrested cell migration. Collectively, these results demonstrate the utility of the 2.5D culture platform to advance our understanding of the effects of stiffness gradients and mechanotransductive signaling on adhesion-independent cell migration.
authors.affiliation: Department of Bioengineering, Santa Clara University, Santa Clara, California, United States of America.
authors.forename: Mark-Phillip
authors.initials: MP
authors.lastname: Pebworth
authors.affiliation: Department of Bioengineering, Santa Clara University, Santa Clara, California, United States of America.
authors.forename: Sabrina A
authors.initials: SA
authors.lastname: Cismas
authors.affiliation: Department of Bioengineering, Santa Clara University, Santa Clara, California, United States of America.
authors.forename: Prashanth
authors.initials: P
authors.lastname: Asuri
mesh_terms: D000464:Alginates; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D024462:Myosin Type II / Q000378:metabolism; D013499:Surface Properties; D054460:rho-Associated Kinases / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D054460:rho-Associated Kinases; D024462:Myosin Type II; D020742:rhoA GTP-Binding Protein