In vitro microphysiological system for high throughput 3D tissue organization and biological function

摘要

Techniques for generating microtissues, including a micro-fabricated platform including at least one micro-well including a plurality of micro-cantilevers coupled thereto and surrounded by a plurality of ridges, each micro-cantilever including a cap at a terminal end thereof. The platform can be immersed in a suspension of cells. The suspension of cells can be driven into at least one micro-well, and the ridges can be de-wetted to remove excess suspension and isolate the suspension of cells in each micro-well. The cells can be driven in the suspension of each micro-well toward a top surface of the suspension, which can be polymerized to form a matrix. The cells can be cultivated to spontaneously compact the matrix such that the micro-cantilevers anchor and constrain the contracting matrix to form a band of microtissue that spans across the micro-cantilevers.

主权项

1. A method for generating microtissues, comprising:
providing a micro-fabricated platform including at least one micro-well including a plurality of micro-cantilevers coupled to the bottom surface of each of at least one micro-well, each micro-cantilever including a cap at a terminal end thereof, wherein each of at least one micro-well is surrounded by a plurality of ridges; immersing the platform in a suspension of cells; driving the suspension of cells into the at least one micro-well; de-wetting the ridges to remove excess suspension, thereby isolating the suspension of cells in each of at least one micro-well; polymerizing the suspension to form a matrix; cultivating the cells over time, to spontaneously compact the matrix, wherein the micro-cantilevers anchor the contracting matrix, constraining the contraction of the matrix to form a band of microtissue that spans across the micro-cantilevers; and measuring a contractile function of the band of microtissue, wherein measuring comprises: imaging the micro-fabricated platform, over time, to acquire image data determining a force exerted on the micro-cantilevers based on at least the image data and a spring constant corresponding to each micro-cantilever; and identifying a contractile function of the band of microtissue based on at least the determined force, wherein the contractile function includes one or more of beat frequency, contraction duration, change in beat frequency over time, change in contraction duration over time, or variance in any of these parameters.