Nature Methods2, 119 - 125 (2005)
Published online: 21 January 2005; | doi:10.1038/nmeth736
An extracellular matrix microarray for probing cellular differentiation
Christopher J Flaim, Shu Chien
& Sangeeta N Bhatia
Departments of Bioengineering and Medicine, University of California San Diego, 9500 Gilman Drive- MC 0412, La Jolla, California 92093-0412, USA.
Correspondence should be addressed to Sangeeta N Bhatia sbhatia@ucsd.edu
We present an extracellular matrix (ECM) microarray platform for the culture of patterned cells atop combinatorial matrix mixtures. This platform enables the study of differentiation in response to a multitude of microenvironments in parallel. The fabrication process required only access to a standard robotic DNA spotter, off-the-shelf materials and 1,000 times less protein than conventional means of investigating cell-ECM interactions. To demonstrate its utility, we applied this platform to study the effects of 32 different combinations of five extracellular matrix molecules (collagen I, collagen III, collagen IV, laminin and fibronectin) on cellular differentiation in two contexts: maintenance of primary rat hepatocyte phenotype indicated by intracellular albumin staining and differentiation of mouse embryonic stem (ES) cells toward an early hepatic fate, indicated by expression of a -galactosidase reporter fused to the fetal liver-specific gene, Ankrd17 (also known as gtar). Using this technique, we identified combinations of ECM that synergistically impacted both hepatocyte function and ES cell differentiation. This versatile technique can be easily adapted to other applications, as it is amenable to studying almost any insoluble microenvironmental cue in a combinatorial fashion and is compatible with several cell types.
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated.
Natureproducts is an online service detailing information about specific
products used in this article, you can view the product descriptions, request
information and compare with other similar products. The products
used are listed in alphabetical order.
-galactosidase reporter fused to the fetal liver-specific gene, Ankrd17 (also known as gtar). Using this technique, we identified combinations of ECM that synergistically impacted both hepatocyte function and ES cell differentiation. This versatile technique can be easily adapted to other applications, as it is amenable to studying almost any insoluble microenvironmental cue in a combinatorial fashion and is compatible with several cell types.
