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Patterning of Fibroblast and Matrix Anisotropy within 3D Confinement is Driven by the Cytoskeleton
被引:11
作者:
Serbo, Janna V.
[1
]
Kuo, Scot
[1
,2
]
Lewis, Shawna
[3
]
Lehmann, Matthew
[4
]
Li, Jiuru
[4
]
Gracias, David H.
[4
,5
]
Romer, Lewis H.
[1
,2
,3
,6
,7
]
机构:
[1] Johns Hopkins Univ, Sch Med, Biomed Engn, Baltimore, MD 21287 USA
[2] Johns Hopkins Univ, Sch Med, Cell Biol, Baltimore, MD 21287 USA
[3] Johns Hopkins Univ, Sch Med, Anesthesiol & Crit Care Med, Baltimore, MD 21287 USA
[4] Johns Hopkins Univ, Chem & Biomol Engn, Baltimore, MD 21218 USA
[5] Johns Hopkins Univ, Mat Sci & Engn, Baltimore, MD 21218 USA
[6] Johns Hopkins Univ, Sch Med, Pediat, Baltimore, MD 21287 USA
[7] Johns Hopkins Univ, Sch Med, Ctr Cell Dynam, Baltimore, MD 21287 USA
基金:
美国国家科学基金会;
关键词:
EXTRACELLULAR-MATRIX;
CELL-MIGRATION;
DEPOLYMERIZATION;
ALIGNMENT;
PATHWAYS;
COMPLEX;
TENSION;
GROWTH;
SHAPE;
D O I:
10.1002/adhm.201500030
中图分类号:
R318 [生物医学工程];
学科分类号:
0831 ;
摘要:
Effects of 3D confinement on cellular growth and matrix assembly are important in tissue engineering, developmental biology, and regenerative medicine. Polydimethylsiloxane wells with varying anisotropy are microfabicated using soft-lithography. Microcontact printing of bovine serum albumin is used to block cell adhesion to surfaces between wells. The orientations of fibroblast stress fibers, microtubules, and fibronectin fibrils are examined 1 day after cell seeding using laser scanning confocal microscopy, and anisotropy is quantified using a custom autocorrelation analysis. Actin, microtubules, and fibronectin exhibit higher anisotropy coefficients for cells grown in rectangular wells with aspect ratios of 1: 4 and 1: 8, as compared to those in wells with lower aspect ratios or in square wells. The effects of disabling individual cytoskeletal components on fibroblast responses to anisotropy are then tested by applying actin or microtubule polymerization inhibitors, Rho kinase inhibitor, or by siRNA-mediated knockdown of AXL or cofilin-1. Latrunculin A decreases cytoskeletal and matrix anisotropy, nocodazole ablates both, and Y27632 mutes cellular polarity while decreasing matrix anisotropy. AXL siRNA knockdown has little effect, as does siRNA knockdown of cofilin-1. These data identify several specific cytoskeletal strategies as targets for the manipulation of anisotropy in 3D tissue constructs.
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页码:146 / 158
页数:13
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