Patterning of cell-instructive hydrogels by hydrodynamic flow focusing

Cosson, Steffen, Allazetta, Simone and Lutolf, Matthias P. (2013) Patterning of cell-instructive hydrogels by hydrodynamic flow focusing. Lab on a Chip, 13 11: 2099-2105. doi:10.1039/c3lc50219h

Author Cosson, Steffen
Allazetta, Simone
Lutolf, Matthias P.
Title Patterning of cell-instructive hydrogels by hydrodynamic flow focusing
Journal name Lab on a Chip   Check publisher's open access policy
ISSN 1473-0197
Publication date 2013-01-01
Sub-type Article (original research)
DOI 10.1039/c3lc50219h
Open Access Status Not yet assessed
Volume 13
Issue 11
Start page 2099
End page 2105
Total pages 7
Place of publication Cambridge, United Kingdom
Publisher R S C Publications
Language eng
Abstract Microfluidic gradient systems offer a very precise means to probe the response of cells to graded biomolecular signals in vitro, for example to model how morphogen proteins affect cell fate during developmental processes. However, existing gradient makers are designed for non-physiological plastic or glass cell culture substrates that are often limited in maintaining the phenotype and function of difficult-to-culture mammalian cell types, such as stem cells. To address this bottleneck, we combine hydrogel engineering and microfluidics to generate tethered protein gradients on the surface of biomimetic poly(ethylene glycol) (PEG) hydrogels. Here we used software-assisted hydrodynamic flow focusing for exposing and rapidly capturing tagged proteins to gels in a step-wise fashion, resulting in immobilized gradients of virtually any desired shape and composition. To render our strategy amenable for high-throughput screening of multifactorial artificial cellular microenvironments, a dedicated microfluidic chip was devised for parallelization and multiplexing, yielding arrays of orthogonally overlapping gradients of up to 4 × 4 proteins. To illustrate the power of the platform for stem cell biology, we assessed how gradients of tethered leukemia inhibitory factor (LIF) influence embryonic stem cell (ESC) behavior. ESC responded to LIF gradients in a binary manner, maintaining the pluripotency marker Rex1/Zfp42 and forming self-renewing colonies above a threshold concentration of 85 ng cm-2. Our concept should be broadly applicable to probe how complex signaling microenvironments influence stem cell fate in culture.
Keyword Morphogen gradient
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
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Citation counts: TR Web of Science Citation Count  Cited 14 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 18 times in Scopus Article | Citations
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Created: Fri, 08 Aug 2014, 20:32:14 EST by Steffen Cosson on behalf of Aust Institute for Bioengineering & Nanotechnology