title: Experimental models for investigating intra-stromal migration of corneal keratocytes, fibroblasts and myofibroblasts.
abstract: Following laser vision correction, corneal keratocytes must repopulate areas of cell loss by migrating through the intact corneal stroma, and this can impact corneal shape and transparency. In this study, we evaluate 3D culture models for simulating this process in vitro. Buttons (8 mm diameter) were first punched out of keratocyte populated compressed collagen matrices, exposed to a 3mm diameter freeze injury, and cultured in serum-free media (basal media) or media supplemented with 10% FBS, TGFβ1 or PDGF BB. Following freeze injury, a region of cell death was observed in the center of the constructs. Although cells readily migrated on top of the matrices to cover the wound area, a limited amount of cell migration was observed within the constructs. We next developed a novel "sandwich" model, which better mimics the native lamellar architecture of the cornea. Using this model, significant migration was observed under all conditions studied. In both models, cells in TGFβ and 10% FBS developed stress fibers; whereas cells in PDGF were more dendritic. PDGF stimulated the most inter-lamellar migration in the sandwich construct. Overall, these models provide insights into the complex interplay between growth factors, cell mechanical phenotypes and the structural properties of the ECM.
authors.affiliation: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, TX; matthew.petroll@utsouthwestern.edu (W.P.); neema.lakshman@utsouthwestern.edu (N.L.); lisha.ma@utsouthwestern.edu (L.M.).
authors.forename: W Matthew
authors.initials: WM
authors.lastname: Petroll
authors.forename: Neema
authors.initials: N
authors.lastname: Lakshman
authors.forename: Lisha
authors.initials: L
authors.lastname: Ma
keywords: 3D Culture; Cell Mechanics; Corneal Keratocytes; Extracellular Matrix; Growth Factors