Differentiated cells make up tissues and organs, and communicate within a complex, three dimensional (3D) environment. The spatial arrangement of cellular interactions is difficult to recapitulate in vitro. Here, a simple and rapid method for stepwise formation of 2D multicellular structures through the biotin‐streptavidin (SA) interaction and further construction of controlled, 3D, multilayered, tissue‐like structures by using the stress‐induced rolling membrane (SIRM) technique is reported. The biotinylated cells connect with the SA‐coated adherent cells to form a bilayer. The bilayer of two types of cells on the SIRM is transformed into 3D tubes, in which two types of cells can directly interact and communicate with each other, mimicking the in vivo conditions of tubular structures such as blood vessel. This method has the potential to recapitulate functional tubular structures for tissue engineering.

We present a device for stretching cells adhering to elastic membranes in equiaxial or uniaxial mode, meanwhile allowing real-time imaging of molecular dynamics of live cells at high resolution on an inverted microscope during the entire process of the stretch. We obtained high-resolution images of stress fibers at each stage of the stretch, and found that stress fibers were shortened after one stretching cycle. We, for the first time, captured real-time images of the process of stress fiber disassembly during stretching. Several adjacent stress fibers appeared to reassemble into a single one after stretching. All these indicated that mechanical stretching played important roles in the rearrangement of actin filaments. This device will be especially useful in studies of the molecular dynamics in the process of mechanotransduction. The device is fabricated on a glass slide through a simple procedure and is adaptable to most ordinary laboratories.

Different tissues have specific mechanical properties and cells of different geometries, such as elongated muscle cells and polygonal endothelial cells, which are precisely regulated during embryo development. However, the mechanisms that underlie these processes are not clear. Here, we built an in vitro model to mimic the cellular microenvironment of muscle by combining both mechanical stretch and geometrical control. We found that mechanical stretch was a key factor that determined the optimal geometry of myoblast C2C12 cells under stretch, whereas vascular endothelial cells and fibroblasts had no such dependency. We presented the first experimental evidence that can explain why myoblasts are destined to take the elongated geometry so as to survive and maintain parallel actin filaments along the stretching direction. The study is not only meaningful for the research on myogenesis but also has potential application in regenerative medicine.

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