Mechanoactivation of Single Stem Cells in Microgels Using a 3D‐Printed Stimulation Device

Abstract

In this study, the novel 3D‐printed pressure chamber for encapsulated single‐cell stimulation (3D‐PRESS) platform is introduced for the mechanical stimulation of single stem cells in 3D microgels. The custom‐designed 3D‐PRESS, allows precise pressure application up to 400 kPa at the single‐cell level. Microfluidics is employed to encapsulate single mesenchymal stem cells within ionically cross‐linked alginate microgels with cell adhesion RGD peptides. Rigorous testing affirms the leak‐proof performance of the 3D‐PRESS device up to 400 kPa, which is fully biocompatible. 3D‐PRESS is implemented on mesenchymal stem cells for mechanotransduction studies, by specifically targeting intracellular calcium signaling and the nuclear translocation of a mechanically sensitive transcription factor. Applying 200 kPa pressure on individually encapsulated stem cells reveals heightened calcium signaling in 3D microgels compared to conventional 2D culture. Similarly, Yes‐associated protein (YAP) translocation into the nucleus occurs at 200 kPa in 3D microgels with cell‐binding RGD peptides unveiling the involvement of integrin‐mediated mechanotransduction in singly encapsulated stem cells in 3D microgels. Combining live‐cell imaging with precise mechanical control, the 3D‐PRESS platform emerges as a versatile tool for exploring cellular responses to pressure stimuli, applicable to various cell types, providing novel insights into single‐cell mechanobiology.