3D ECM-rich environment sustains the identity of naive human iPSCs
Authors listElisa Cesare Anna Urciuolo Hannah Stuart Erika Torchio Alessia Gesualdo Cecilia Laterza Onelia Gagliano Sebastian Martewicz Meihua Cui Anna Manfredi Lucio Di Filippo Patrizia Sabatelli Stefano Squarzoni Irene Zorzan Riccardo M Betto Graziano Martello Davide Cacchiarelli Camilla Luni Nicola Elvassore
The establishment of in vitro naive human pluripotent stem cell cultures opened new perspectives for the study of early events in human development. The role of several transcription factors and signaling pathways have been characterized during maintenance of human naive pluripotency. However, little is known about the role exerted by the extracellular matrix (ECM) and its three-dimensional (3D) organization. Here, using an unbiased and integrated approach combining microfluidic cultures with transcriptional, proteomic, and secretome analyses, we found that naive, but not primed, hiPSC colonies are characterized by a self-organized ECM-rich microenvironment. Based on this, we developed a 3D culture system that supports robust long-term feeder-free self-renewal of naive hiPSCs and also allows direct and timely developmental morphogenesis simply by modulating the signaling environment. Our study opens new perspectives for future applications of naive hiPSCs to study critical stages of human development in 3D starting from a single cell.
Journal Cell Stem Cell
Issue number 12