An extracellular matrix microarray for probing cellular differentiation.

Publication Type:

Journal Article

Source:

Nat Methods, Volume 2, Issue 2, p.119-25 (2005)

Keywords:

Animals, Cell Culture Techniques, Cell Differentiation, Cell Line, Combinatorial Chemistry Techniques, Equipment Design, Equipment Failure Analysis, Extracellular Matrix Proteins, Hepatocytes, Mice, Microarray Analysis, Microfluidic Analytical Techniques, Molecular Probe Techniques, Rats, Robotics, Stem Cells

Abstract:

<p>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 beta-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.</p>

PDF: