HMEC-1 and iMSC#3 cells were cultured on surfaces coated with thermoresponsive polymers, which allow a temperature-triggered cell detachment, keeping the cells within their extracellular matrix. The polymer coating enables cell adhesion during culture at 37 °C as well as cell detachment when the grown cell sheet is exposed to lower temperatures. After detachment, the cell sheets were rolled onto a coated rod and fluorescently stained for microscopy. The video shows images of a z-stack of rolled HMEC-1 (inner layers) and iMSC#3 (outer layers) cell sheets. F-actin staining is depicted in yellow. The vertical alignment of iMSC#3 cells can be observed and the inner layer of HMEC-1 cells can be identified by cobblestone morphology.