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Issue 12, 2012
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Responsive culture platform to examine the influence of microenvironmental geometry on cell function in 3D

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Abstract

We describe the development of a well-based cell culture platform that enables experimenters to control the geometry and connectivity of cellular microenvironments spatiotemporally. The base material is a hydrogel comprised of photolabile and enzyme-labile crosslinks and pendant cell adhesion sequences, enabling spatially-specific, in situ patterning with light and cell-dictated microenvironment remodeling through enzyme secretion. Arrays of culture wells of varying shape and size were patterned into the hydrogel surface using photolithography, where well depth was correlated with irradiation dose. The geometry of these devices can be subsequently modified through sequential patterning, while simultaneously monitoring changes in cell geometry and connectivity. Towards establishing the utility of these devices for dynamic evaluation of the influence of physical cues on tissue morphogenesis, the effect of well shape on lung epithelial cell differentiation (i.e., primary mouse alveolar type II cells, ATII cells) was assessed. Shapes inspired by alveoli were degraded into hydrogel surfaces. ATII cells were seeded within the well-based arrays and encapsulated by the addition of a top hydrogel layer. Cell differentiation in response to these geometries was characterized over 7 days of culture with immunocytochemistry (surfactant protein C, ATII; T1α protein, alveolar type I (ATI) differentiated epithelial cells) and confocal image analysis. Individual cell clusters were further connected by eroding channels between wells during culture via controlled two-photon irradiation. Collectively, these studies demonstrate the development and utility of responsive hydrogel culture devices to study how a range of microenvironment geometries of evolving shape and connectivity might influence or direct cell function.

Graphical abstract: Responsive culture platform to examine the influence of microenvironmental geometry on cell function in 3D

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Publication details

The article was received on 05 Sep 2012, accepted on 27 Oct 2012 and first published on 31 Oct 2012


Article type: Technical Innovation
DOI: 10.1039/C2IB20212C
Citation: Integr. Biol., 2012,4, 1540-1549
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    Responsive culture platform to examine the influence of microenvironmental geometry on cell function in 3D

    A. M. Kloxin, K. J. R. Lewis, C. A. DeForest, G. Seedorf, M. W. Tibbitt, V. Balasubramaniam and K. S. Anseth, Integr. Biol., 2012, 4, 1540
    DOI: 10.1039/C2IB20212C

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