3D multicellular culture systems for real time insights in physical and chemical signals underlying stem cell morphogenesis and lineage fateAcronym: 3DMuSyC

Although many studies have been able to create differentiated progeny from pluripotent stem cells (PSC) using conventional 2D cultures, these approaches cannot recreate the complex morphogenesis and phenotypic composition of in-vivo tissues and organs. Therefore, we will develop advanced 3D multicellular cell culture systems to investigate the interaction between cells and their microenvironment (cell-cell interactions and interactions with physical and chemical properties of the extracellular matrix, ECM) that support and help develop their fate and assembly into organized tissue-like constructs. Here, we propose to develop and integrate several innovative technology platforms which can (1) define the appropriate molecular (ECM content) and biophysical (ECM physical properties) cues, and soluble factors (morphogens and small molecules) in the context of differentiation and morphogenesis, that will (2) allow real time assessment of cell fate changes and cell-cell-ECM interactions within 3D cultures using smart fluorophores (3) incorporated by CRISPR/Cas and eFlut technology in PSCs and (4) wherein mechanical forces at the cellular and subcellular scale can be probed. Such advanced 3D cultures should enable a greater understanding of early morphogenesis and subsequent fating to skeletal muscle, heart or liver from stem cells, at the single cell as well as multicellular level, including the role of external chemical and physical cues. We believe that this highly innovative and challenging proposal will be feasible by the joining of forces of a highly interdisciplinary team of young and senior investigators with expertise in stem cell biology, bioengineering, biosensor chemistry and mechanobiology from the Department of Development and Regeneration, within the Group Biomedical Sciences and the Departments of Biochemistry, Molecular and Structural Biology and of Biomechanics, within the Group Science, Engineering and Technology. This unique combination of expertise will allow us for the first time to explore the role of the microenvironment on multicellular tissue-like constructs in a dynamic, real-time and systematic manner. We expect the technologies developed and the insights gained in this project to be broadly applicable to a better understanding of 3D morphogenesis in the context of in-vitro developmental biology, for stem-cell derived regenerative medicine applications as well as for disease modelling.

Keywords:stem cell

Disciplines:Laboratory medicine, Palliative care and end-of-life care, Regenerative medicine, Other basic sciences, Other health sciences, Nursing, Other paramedical sciences, Other translational sciences, Other medical and health sciences