Facilitated Expansion of Human Embryonic Stem Cells by Single-Cell Enzymatic Dissociation

摘要: Traditionally, human embryonic stem cells (hESCs) are propagated by mechanical dissection or enzymatic dissociation into clusters of cells. To facilitate up-scaling and the use hESC in various experimental manipulations, such as fluorescence-activated cell sorting, electroporation, clonal selection, it is important to develop new, stable culture systems based on single-cell propagation. Here, we show that hESCs, which were derived passaged dissection, can be rapidly adjusted propagation single As an indication stability this system, demonstrate hESCs maintained undifferentiated, pluripotent, genetically normal state for up 40 passages. We also a recombinant trypsin preparation increases survival compared with porcine trypsin. Finally, foreskin fibroblast feeders superior commonly used mouse terms their ability prevent spontaneous differentiation after passaging. Importantly, system widely applicable should therefore general reliable large-scale cultivation well manipulations.