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.
引用
收藏
页码:146 / 158
页数:13
相关论文
共 55 条
[1]   Scalable alignment of three-dimensional cellular constructs in a microfluidic chip [J].
Anene-Nzelu, Chukwuemeka George ;
Peh, Kah Yim ;
Fraiszudeen, Azmall ;
Kuan, Yee Han ;
Ng, Sum Huan ;
Toh, Yi Chin ;
Leo, Hwa Liang ;
Yu, Hanry .
LAB ON A CHIP, 2013, 13 (20) :4124-4133
[2]   Directed 3D cell alignment and elongation in microengineered hydrogels [J].
Aubin, Hug ;
Nichol, Jason W. ;
Hutson, Che B. ;
Bae, Hojae ;
Sieminski, Alisha L. ;
Cropek, Donald M. ;
Akhyari, Payam ;
Khademhosseini, Ali .
BIOMATERIALS, 2010, 31 (27) :6941-6951
[3]   Stem Cell Sources for Vascular Tissue Engineering and Regeneration [J].
Bajpai, Vivek K. ;
Andreadis, Stelios T. .
TISSUE ENGINEERING PART B-REVIEWS, 2012, 18 (05) :405-425
[4]   Deconstructing the third dimension - how 3D culture microenvironments alter cellular cues [J].
Baker, Brendon M. ;
Chen, Christopher S. .
JOURNAL OF CELL SCIENCE, 2012, 125 (13) :3015-3024
[5]   From Single Cells to Tissues: Interactions between the Matrix and Human Breast Cells in Real Time [J].
Barnes, Clifford ;
Speroni, Lucia ;
Quinn, Kyle P. ;
Montevil, Mael ;
Saetzler, Kurt ;
Bode-Animashaun, Gbemisola ;
McKerr, George ;
Georgakoudi, Irene ;
Downes, C. Stephen ;
Sonnenschein, Carlos ;
Howard, C. Vyvyan ;
Soto, Ana M. .
PLOS ONE, 2014, 9 (04)
[6]   Controlling the structural and functional anisotropy of engineered cardiac tissues [J].
Bian, Weining ;
Jackman, Christopher P. ;
Bursac, Nenad .
BIOFABRICATION, 2014, 6 (02)
[7]   Endothelial Matrix Assembly during Capillary Morphogenesis: Insights from Chimeric TagRFP-Fibronectin Matrix [J].
Chang, Fumin ;
Lemmon, Christopher A. ;
Nilaratanakul, Voraphoj ;
Rotter, Varda ;
Romer, Lewis .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 2014, 62 (11) :774-790
[8]   Geometric control of cell life and death [J].
Chen, CS ;
Mrksich, M ;
Huang, S ;
Whitesides, GM ;
Ingber, DE .
SCIENCE, 1997, 276 (5317) :1425-1428
[9]   Rho-stimulated contractility drives the formation of stress fibers and focal adhesions [J].
ChrzanowskaWodnicka, M ;
Burridge, K .
JOURNAL OF CELL BIOLOGY, 1996, 133 (06) :1403-1415
[10]   INHIBITION OF ACTIN POLYMERIZATION BY LATRUNCULIN-A [J].
COUE, M ;
BRENNER, SL ;
SPECTOR, I ;
KORN, ED .
FEBS LETTERS, 1987, 213 (02) :316-318