A low-cost uniaxial cell stretcher for six parallel wells
Abstract: Cells in the lungs, the heart, and numerous other organs, are constantly exposed to dynamic forces and deformations. To mimic these dynamic mechanical loading conditions and to study the resulting cellular responses such as morphological changes or the activation of biochemical signaling pathways, cells are typically seeded on flexible 2D substrates that are uniaxially or biaxially stretched. Here, we present an open-source cell stretcher built from parts of an Anet A8 3D printer. The cell stretcher is controlled by a fully programmable open-source software using GCode and Python. Up to 6 flexible optically clear substrates can be stretched simultaneously, allowing for comparative multi-batch biological studies including microscopic image analysis. The cell yield from the cell culture area of 4 cm2 per dish is sufficient for Western-blot protein analysis. As a proof-of-concept, we study the activation of the Yes-associated protein (YAP) mechanotransduction pathway in response to increased cytoskeletal tension induced by uniaxial stretching of epithelial cells. Our data support the previously observed activation of the YAP transcription pathway by stretch-induced increase of cytoskeletal tension and demonstrate the suitability of the cell stretcher to study complex mechano-biological processes. ----- Files: • Acryl3MM.pdf: Ready-to-cut file for the acrylic glass parts needed to build the electronics and controller unit housing and the cell substrate molds. • Acryl6MM.pdf: Ready-to-cut file for the acrylic glass parts needed to build the stretcher unit. • MoldFrame.stl: Ready-to-print file for the 3D printed mold frame. • Clamp.stl: Ready-to-print file for the 3D printed stretching clamps. • Stretcher.zip: Customized version of the Raspbian operating version for the Raspberry Pi. • Raspberry_Pi_Imager_v1.2.zip: Open-source software for Windows PCs to install the Raspberry Pi operating system on a memory card. • Skynet_for_Stretcher.zip: Modified version of the SkyNet3D (Marlin) open-source firmware for Anet 3D printers. • Arduino-1.8.0.zip: Modified version of the Arduino integrated development environment for Windows PCs with a pre-installed driver for uploading software to the ANET A8 mainboard.