TY - JOUR
T1 - Corneal stromal cells use both high- and low-contractility migration mechanisms in 3-D collagen matrices
AU - Kim, Areum
AU - Zhou, Chengxin
AU - Lakshman, Neema
AU - Petroll, W. Matthew
N1 - Funding Information:
Supported by NIH R01 EY013322 , NIH P30 EY020799 , and Research to Prevent Blindness, Inc .
Funding Information:
This study was supported in part by NIH R01 EY 013322 , NIH P30 EY020799 , and an unrestricted grant and Senior Scientific Investigator Award (WMP) from Research to Prevent Blindness, Inc., NY, NY . The authors wish to thank Dr. Dwight Cavanagh for his helpful comments and suggestions.
PY - 2012/4/1
Y1 - 2012/4/1
N2 - Corneal keratocyte migration can impact both corneal clarity and refractive outcome following injury or refractive surgery. In this study, we investigated how culture conditions, ECM properties, and Rho kinase activity regulate the mechanics of keratocyte migration, using a nested collagen matrix model. Time-lapse imaging demonstrated that both serum and PDGF stimulate keratocyte migration into the outer matrix. Although the velocity of cell migration was similar, cells in serum were bipolar and induced significant matrix deformation during migration, whereas PDGF induced extension of branching dendritic processes with smaller, more localized force generation. These differences in cell-induced matrix reorganization were verified with a global matrix contraction assay and confocal reflection imaging, using both bovine and rat tail collagen. When constructs were detached from the substrate to lower the effective stiffness, migration was significantly reduced in serum; but was unchanged in PDGF. These differences in migration mechanics were mediated, in part, by Rho kinase. Overall, corneal keratocytes can effectively migrate through collagen matrices using varying degrees of cellular force generation. Low-contractility migration may facilitate keratocyte repopulation of the stroma following surgery or injury, without altering the structural and mechanical properties that are critical to maintaining corneal transparency.
AB - Corneal keratocyte migration can impact both corneal clarity and refractive outcome following injury or refractive surgery. In this study, we investigated how culture conditions, ECM properties, and Rho kinase activity regulate the mechanics of keratocyte migration, using a nested collagen matrix model. Time-lapse imaging demonstrated that both serum and PDGF stimulate keratocyte migration into the outer matrix. Although the velocity of cell migration was similar, cells in serum were bipolar and induced significant matrix deformation during migration, whereas PDGF induced extension of branching dendritic processes with smaller, more localized force generation. These differences in cell-induced matrix reorganization were verified with a global matrix contraction assay and confocal reflection imaging, using both bovine and rat tail collagen. When constructs were detached from the substrate to lower the effective stiffness, migration was significantly reduced in serum; but was unchanged in PDGF. These differences in migration mechanics were mediated, in part, by Rho kinase. Overall, corneal keratocytes can effectively migrate through collagen matrices using varying degrees of cellular force generation. Low-contractility migration may facilitate keratocyte repopulation of the stroma following surgery or injury, without altering the structural and mechanical properties that are critical to maintaining corneal transparency.
KW - Cell mechanics
KW - Cell migration
KW - Corneal keratocytes
KW - Rho kinase
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U2 - 10.1016/j.yexcr.2011.12.018
DO - 10.1016/j.yexcr.2011.12.018
M3 - Article
C2 - 22233682
AN - SCOPUS:84857656057
SN - 0014-4827
VL - 318
SP - 741
EP - 752
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 6
ER -