Reprogramming Shape

By Timothée Vignaud, Qingzong Tseng, Manuel Théry, iRTSV in Grenoble, France

Micropatterning can also control a cell’s size and shape. Here Thery and colleagues apply adhesive molecules (e.g., fibronectin) on glass slides in varies shapes—a “T” (top right) or an “H” (bottom right). When they plate one or two cells onto the micropattern, they adopts a convex “envelop” shape around the whole micropattern: the single cell becomes triangular on a T pattern and the cell doublet forms a square on the H. If they “draw” a micropattern near a fixed cell on the plate (left), the cell progressively spreads on this new bar and assembles stress fibers attached to its extremity

Left: A RPE1 cell expresses LifeAct-GFP, which labels the actin network in living cells. After the micropattern is drawn near the cell, images were acquired every 20 minutes with an inverted TE2000 Nikon microscope (100x oil objective). Colors arbitrarily designate each stage of reprogramming.

Right: Single RPE1 cells on the T (top) and an MCF10A cell doublet on the H (bottom) were permeabilized and fixed with paraformaldehyde after plated on the micropatterned glass slide. The actin network and focal adhesions are labeled green (phaloidin-FITC) and red (antibodies against vinculin or paxilin), respectively, while intercellular junctions are labeled white with antibodies agaisnt &bega;-catenin. Images acquired with a Leica DMRA microscope (100X oil objective).


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