Spatially controlled stem cell differentiation via morphogen gradients: A comparison of static and dynamic microfluidic platforms

Abstract

The ability to harness the processes by which complex tissues arise during embryonic development would improve the ability to engineer complex tissuelike constructs in vitro—a longstanding goal of tissue engineering and regenerative medicine. In embryos, uniform populations of stem cells are exposed to spatial gradients of diffusible extracellular signaling proteins, known as morphogens. Varying levels of these signaling proteins induce stem cells to differentiate into distinct cell types at different positions along the gradient, thus creating spatially patterned tissues. Here, the authors describe two straightforward and easy-to-adopt microfluidic strategies to expose human pluripotent stem cells in vitro to spatial gradients of desired differentiation-inducing extracellular signals. Both approaches afford a high degree of control over the distribution of extracellular signals, while preserving the viability of the cultured stem cells. The first microfluidic platform is commercially available and entails static culture, whereas the second microfluidic platform requires fabrication and dynamic fluid exchange. In each platform, the authors first computationally modeled the spatial distribution of differentiation-inducing extracellular signals. Then, the authors used each platform to expose human pluripotent stem cells to a gradient of these signals (in this case, inducing a cell type known as the primitive streak), resulting in a regionalized culture with differentiated primitive streak cells predominately localized on one side and undifferentiated stem cells at the other side of the device. By combining this approach with a fluorescent reporter for differentiated cells and live-cell fluorescence imaging, the authors characterized the spatial and temporal dynamics of primitive streak differentiation within the induced signaling gradients. Microfluidic approaches to create precisely controlled morphogen gradients will add to the stem cell and developmental biology toolkit, and may eventually pave the way to create increasingly spatially patterned tissuelike constructs in vitro.

Microengineered systems with iPSC-derived cardiac and hepatic cells to evaluate drug adverse effects

“…208 Additionally, microfluidic systems can be designed to create signaling gradients within a culture to spatially influence cell response or development. For example, Cui et al 209 demonstrated regionalized formation of the primitive streak from pluripotent stem cells within a device using controlled microfluidics. This same approach could be applied to developmental models to optimize differentiation or maturation conditions, differentiate co-cultures within a device, or for downstream evaluation of various drug concentration gradients.…”

Digital light processing in a hybrid atomic force microscope: In Situ, nanoscale characterization of the printing process

“…Three dimensional (3D) printing, or additive manufacturing (AM), is touted as the next generation of agile, efficient manufacturing technology with the ability to fabricate complex structures for applications ranging from inexpensive rapid prototyping to tissue engineering for regenerative medicine. [1][2][3] Unfortunately, most AM processes introduce micrometer-scale anisotropic inhomogeneities in chemical, thermal, and mechanical properties, causing the performance of fabricated parts to depend strongly and unpredictably on printing conditions. [4,5] Without full understanding of how AM parameters affect mechanical properties, AM will have limited societal impact.…”