Nat Biotechnol. 2010 Apr;28(4):361-4. doi: 10.1038/nbt.1616. Epub 2010 Mar 28.
Undifferentiated human embryonic stem cells (hESCs) are currently propagated on a relatively small scale as monolayer colonies. Culture of hESCs as floating aggregates is widely used for induction of differentiation into embryoid bodies. Here we show that hESC lines can be derived from floating inner cell masses in suspension culture conditions that do not involve feeder cells or microcarriers. This culture system supports prolonged propagation of the pluripotent stem cells as floating clusters without their differentiation into embryoid bodies. HESCs cultivated as aggregates in suspension maintain the expression of pluripotency markers and can differentiate into progeny of the three germ layers both in vitro and in vivo. We further show the controlled differentiation of hESC clusters in suspension into neural spheres. These results pave the way for large-scale expansion and controlled differentiation of hESCs in suspension, which would be valuable in basic and applied research.
未分化的人类胚胎干细胞（hESCs）目前以单层集落的形式进行相对小规模的扩增。将 hESC 培养为悬浮聚集物广泛用于诱导其分化为胚状体。在这里，我们展示了可以从悬浮培养条件下的悬浮内细胞团中衍生出 hESC 系，而无需饲养细胞或微载体。该培养系统支持多能干细胞作为悬浮集落进行长时间扩增，而不会分化为胚状体。在悬浮状态下培养的 hESC 集落作为聚集物保持多能性标志物的表达，并且可以在体外和体内分化为三个胚层的后代。我们进一步展示了在悬浮状态下 hESC 集落的受控分化为神经球体。这些结果为 hESC 在悬浮状态下的大规模扩增和受控分化铺平了道路，这在基础和应用研究中是有价值的。
