Design, fabrication and implementation of a novel multi-parameter control microfluidic platform for three-dimensional cell culture and real-time imaging

被引:276
作者
Vickerman, Vernella [2 ]
Blundo, Jennifer [4 ]
Chung, Seok [1 ]
Kamm, Roger [1 ,3 ]
机构
[1] MIT, Dept Biol Engn, Cambridge, MA 02139 USA
[2] MIT, Dept Chem Engn, Cambridge, MA 02139 USA
[3] MIT, Dept Mech Engn, Cambridge, MA 02139 USA
[4] Stanford Univ, Dept Mech Engn, Stanford, CA 94305 USA
关键词
D O I
10.1039/b802395f
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
New and more biologically relevant in vitro models are needed for use in drug development, regenerative medicine, and fundamental scientific investigation. While the importance of the extracellular microenvironment is clear, the ability to investigate the effects of physiologically relevant biophysical and biochemical factors is restricted in traditional cell culture platforms. Moreover, the versatility for multi-parameter manipulation, on a single platform, with the optical resolution to monitor the dynamics of individual cells or small population is lacking. Here we introduce a microfluidic platform for 3D cell culture in biologically derived or synthetic hydrogels with the capability to monitor cellular dynamics in response to changes in their microenvironment. Direct scaffold microinjection, was employed to incorporate 3D matrices into microfluidic devices. Our system geometry permits a unique window for studying directional migration, e.g. sprouting angiogenesis, since sprouts grow predominantly in the microscopic viewing plane. In this study, we demonstrate the ability to generate gradients (non-reactive solute), surface shear, interstitial flow, and image cells in situ. Three different capillary morphogenesis assays are demonstrated. Human adult dermal microvascular endothelial cells (HMVEC-ad) were maintained in culture for up to 7 days during which they formed open lumen-like structures which was confirmed with confocal microscopy and by perfusion with fluorescent microspheres. In the sprouting assay, time-lapse movies revealed cellular mechanisms and dynamics (filopodial projection/retraction, directional migration, cell division and lumen formation) during tip-cell invasion of underlying 3D matrix and subsequent lumen formation.
引用
收藏
页码:1468 / 1477
页数:10
相关论文
共 38 条
[1]   Cell culture: Biology's new dimension [J].
Abbott, A .
NATURE, 2003, 424 (6951) :870-872
[2]   RGD-dependent vacuolation and lumen formation observed during endothelial cell morphogenesis in three-dimensional fibrin matrices involves the αvβ3 and α5β1 integrins [J].
Bayless, KJ ;
Salazar, R ;
Davis, GE .
AMERICAN JOURNAL OF PATHOLOGY, 2000, 156 (05) :1673-1683
[3]   Sphingosine-1-phosphate markedly induces matrix metalloproteinase and integrin-dependent human endothelial cell invasion and lumen formation in three-dimensional collagen and fibrin matrices [J].
Bayless, KJ ;
Davis, GE .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2003, 312 (04) :903-913
[4]   Interstitial flow as a guide for lymphangiogenesis [J].
Boardman, KC ;
Swartz, MA .
CIRCULATION RESEARCH, 2003, 92 (07) :801-808
[5]   Intracellular macromolecular mobility measured by fluorescence recovery after photobleaching with confocal laser scanning microscopes [J].
Braga, J ;
Desterro, JMP ;
Carmo-Fonseca, M .
MOLECULAR BIOLOGY OF THE CELL, 2004, 15 (10) :4749-4760
[6]   Microfabricated platform for studying stem cell fates [J].
Chin, VI ;
Taupin, P ;
Sanga, S ;
Scheel, J ;
Gage, FH ;
Bhatia, SN .
BIOTECHNOLOGY AND BIOENGINEERING, 2004, 88 (03) :399-415
[7]   Patterned deposition of cells and proteins onto surfaces by using three-dimensional microfluidic systems [J].
Chiu, DT ;
Jeon, NL ;
Huang, S ;
Kane, RS ;
Wargo, CJ ;
Choi, IS ;
Ingber, DE ;
Whitesides, GM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (06) :2408-2413
[8]   Molecular mechanisms of blood vessel growth [J].
Conway, EM ;
Collen, D ;
Carmeliet, P .
CARDIOVASCULAR RESEARCH, 2001, 49 (03) :507-521
[9]   Taking cell-matrix adhesions to the third dimension [J].
Cukierman, E ;
Pankov, R ;
Stevens, DR ;
Yamada, KM .
SCIENCE, 2001, 294 (5547) :1708-1712
[10]  
EDDINGTON DTP, 2006, SENSOR ACTUAT B-CHEM, P170