Using 3D Culture to Investigate the Role of Mechanical Signaling in Keratinocyte Stem Cells

摘要: The ability to grow keratinocyte stem cells (KSCs) in 3D culture is an important step forward for investigating the physiological properties of these cells. In epidermis, KSCs are subject various types mechanical stress. To study effects stress on KSCs, monolayer cultures limited as can only form cell-cell contacts one plane and prevent differentiation, grown low (0.05 mM) calcium, which impairs formation calcium-dependent adhesion structures such desmosomes. This contrast how found epidermis vivo, where they connected all sides by other cells, allowing them a more organized cytoskeleton. cytoskeleton essential transducing signals between this cannot be accurately reproduced cultures, do not have same level organization or connections. We describe technique allows generation large numbers uniformly sized cell aggregates using cultured murine KSCs. These produced calcium concentrations (1.2 mM), within with sides, resulting reorganization cytoskeleton, integrating each aggregate. Within aggregates, retain properties, p63 expression, despite increased concentration show activation mitogen-activated protein kinase ERK upon stretch. KSC manipulated further provide physiologically relevant model studying signaling