Mechanochemical actuators of embryonic epithelial contractility.

Spatiotemporal regulation of cell contractility coordinates cell shape change to construct tissue architecture and ultimately directs the morphology and function of the organism. Here we show that contractility responses to spatially and temporally controlled chemical stimuli depend much more strongly on intercellular mechanical connections than on biochemical cues in both stimulated tissues and adjacent cells. We investigate how the cell contractility is triggered within an embryonic epithelial sheet by local ligand stimulation and coordinates a long-range contraction response.

Microscopy tools for quantifying developmental dynamics in Xenopus embryos.

Early Xenopus embryos, and embryonic tissues isolated from them, are excellent model systems to study morphogenesis. Cells migrate, change shape, and differentiate to form new tissues as embryos mature and recapitulate those same processes in tissue isolates. Both large-scale and small-scale cell and tissue movements can be visualized with a range of microscopy techniques.

Epithelial machines of morphogenesis and their potential application in organ assembly and tissue engineering.

Sheets of embryonic epithelial cells coordinate their efforts to create diverse tissue structures such as pits, grooves, tubes, and capsules that lead to organ formation. Such cells can use a number of cell behaviors including contractility, proliferation, and directed movement to create these structures. By contrast, tissue engineers and researchers in regenerative medicine seeking to produce organs for repair or replacement therapy can combine cells with synthetic polymeric scaffolds.

Dynamic control of 3D chemical profiles with a single 2D microfluidic platform.

Dynamic control of three-dimensional (3D) chemical patterns with both high precision and high speed is important in a range of applications from chemical synthesis, flow cytometry, and multi-scale biological manipulation approaches. A central challenge in controlling 3D chemical patterns is the inability to create rapidly tunable 3D profiles with simple and direct approaches that avoid complicated microfabrication. Here, we present the ability to rapidly and precisely create 3D chemical patterns using a single two-dimensional (2D) microfluidic platform.

Detection of dynamic spatiotemporal response to periodic chemical stimulation in a Xenopus embryonic tissue.

Embryonic development is guided by a complex and integrated set of stimuli that results in collective system-wide organization that is both time and space regulated. These regulatory interactions result in the emergence of highly functional units, which are correlated to frequency-modulated stimulation profiles. We have determined the dynamic response of vertebrate embryonic tissues to highly controlled, time-varying localized chemical stimulation using a microfluidic system with feedback control.

Live-cell imaging and quantitative analysis of embryonic epithelial cells in Xenopus laevis.

Embryonic epithelial cells serve as an ideal model to study morphogenesis where multi-cellular tissues undergo changes in their geometry, such as changes in cell surface area and cell height, and where cells undergo mitosis and migrate. Furthermore, epithelial cells can also regulate morphogenetic movements in adjacent tissues(1). A traditional method to study epithelial cells and tissues involve chemical fixation and histological methods to determine cell morphology or localization of particular proteins of interest.

Emergent morphogenesis: elastic mechanics of a self-deforming tissue.

Multicellular organisms are generated by coordinated cell movements during morphogenesis. Convergent extension is a key tissue movement that organizes mesoderm, ectoderm, and endoderm in vertebrate embryos. The goals of researchers studying convergent extension, and morphogenesis in general, include understanding the molecular pathways that control cell identity, establish fields of cell types, and regulate cell behaviors.

Experimental control of excitable embryonic tissues: three stimuli induce rapid epithelial contraction.

Cell generated contractility is a major driver of morphogenesis during processes such as epithelial bending and epithelial-to-mesenchymal transitions. Previous studies of contraction in embryos have relied on developmentally programmed cell shape changes such as those that accompany ventral furrow formation in Drosophila, bottle cell formation in Xenopus, ingression in amniote embryos, and neurulation in vertebrate embryos. We have identified three methods to reproducibly and acutely induce contraction in embryonic epithelial sheets: laser activation, electrical stimulation, and nano-perfusion with chemicals released by wounding.

Variation of cyclic strain parameters regulates development of elastic modulus in fibroblast/substrate constructs.

Dynamic mechanical culture systems are a widely studied approach for improving the functional mechanical properties of tissue engineering constructs intended for loading-bearing orthopedic applications such as tendon/ligament reconstruction. The design of effective mechanical stimulation regimes requires a fundamental understanding of the effects of cyclic strain parameters on the resulting construct properties. Toward this end, these studies employed a modular cyclic strain bioreactor system and fibroblast-seeded, porous polyurethane substrates to systematically investigate the effect of varying cyclic strain amplitude, rate, frequency, and daily cycle number on construct mechanical properties.