A co-culture device with a tunable stiffness to understand combinatorial cell-cell and cell-matrix interactions.
Cell behavior on 2-D in vitro cultures is continually being improved to better mimic in vivo physiological conditions by combining niche cues including multiple cell types and substrate stiffness, which are well known to impact cell phenotype. However, no system exists in which a user can systematically examine cell behavior on a substrate with a specific stiffness (elastic modulus) in culture with a different cell type, while maintaining distinct cell populations. We demonstrate the modification of a silicon reconfigurable co-culture system with a covalently linked hydrogel of user-defined stiffness. This device allows the user to control whether two separate cell populations are in contact with each other or only experience paracrine interactions on substrates of controllable stiffness. To illustrate the utility of this device, we examined the role of substrate stiffness combined with myoblast co-culture on adipose derived stem cell (ASC) differentiation and found that the presence of myoblasts and a 10 kPa substrate stiffness increased ASC myogenesis versus co-culture on stiff substrates. As this example highlights, this technology better controls the in vitro microenvironment, allowing the user to develop a more thorough understanding of the combined effects of cell-cell and cell-matrix interactions.
Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093, USA. christman@eng.ucsd.edu.
Nikhil
N
Rao
Gregory N
GN
Grover
Ludovic G
LG
Vincent
Samantha C
SC
Evans
Yu Suk
YS
Choi
Katrina H
KH
Spencer
Elliot E
EE
Hui
Adam J
AJ
Engler
Karen L
KL
Christman
D000273:Adipose Tissue / Q000166:cytology; D000328:Adult; D002450:Cell Communication; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000295:instrumentation*; D055119:Elastic Modulus; D004867:Equipment Design; D005109:Extracellular Matrix / Q000378:metabolism*; D005782:Gels / Q000737:chemistry; D006801:Humans; D024510:Muscle Development; D032446:Myoblasts / Q000166:cytology*; D011312:Pressure; D012825:Silicon / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology*; D055815:Young Adult
D005782:Gels; D012825:Silicon