Small molecule promoted feeder free and adherent differentiation of functional neurons from human embryonic and induced pluripotent stem cells.
Small molecule promoted feeder free and adherent differentiation of functional neurons from human embryonic and induced pluripotent stem cells.
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发表时间：
2011
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通讯作者：
Danielle N. Drury-Stewart;Ming-ke Song;O. Mohamad;S. Yu;Ling Wei
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作者：
Danielle N. Drury-Stewart;Ming-ke Song;O. Mohamad;S. Yu;Ling Wei
虽然人胚胎干（hES）和诱导多能干（hiPS）细胞在再生医学和发育生物学领域提供了令人兴奋的前景，但这些细胞的有效定向分化仍然很困难。神经诱导方案通常包括悬浮培养或与其他细胞类型共培养，引入异质性并使分析复杂化。此外，昂贵的重组因子通常用于需要数周时间才能完成的过程，使得此类实验在经济上变得困难。我们使用小分子（如dorsomorphin）和常见培养基补充剂开发了一种完全贴壁且无饲养层的神经分化方案。通过巢蛋白染色测量，使用该方案，我们获得了 >90% 的细胞发育成神经前体细胞。源自这些前体的神经元具有电生理活性。经过三周的终末分化，我们获得了在去极化时激发高振幅动作电位的功能性神经元。一部分神经元也会发射重复序列。该协议为涉及文化中神经前体和神经元分化的研究提供了一种更简单且更便宜的方法。
While human embryonic stem (hES) and induced pluripotent stem (hiPS) cells offer exciting prospects in the fields of regenerative medicine and developmental biology, efficient directed differentiation of these cells is still difficult. Neural induction protocols often include suspension culture or co-culture with other cell types, introducing heterogeneity and complicating analysis. In addition, expensive recombinant factors are often used over processes that take weeks to complete, making such experiments financially difficult. We have developed a fully adherent and feeder free neural differentiation protocol using small molecules such as dorsomorphin and common medium supplements. Using this protocol, we obtain >90% of cells developing into neural precursors, as measured by nestin staining. Neurons derived from these precursors are electrophysiologically active. After three weeks of terminal differentiation, we obtain functional neurons which fire high-amplitude action potentials upon depolarization. A subset of neurons also fires repetitive trains. This protocol offers a simpler and less expensive method for investigations involving the differentiation of neural precursors and neurons in culture.