Rapid neurogenesis through transcriptional activation in human stem cell (RNA-Seq)

Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two murine Neurogenin transcription factors in human induced pluripotent stem cells, and obtained neurons with bipolar morphology in four days at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the transition from stem cell to neuron. These profiles were then analyzed to identify the regulatory networks underlying the differentiation of the neurons.

细胞重编程（cellular reprogramming）与干细胞分化（stem cell differentiation）领域的技术进展，现已支持人类神经元分化的体外（ex vivo）研究。然而，阐明其背后的调控程序（regulatory programs）仍颇具挑战：由于神经元分化方案（differentiation protocols）耗时费力，且通常神经元产出率较低。本研究中，我们在人类诱导多能干细胞（human induced pluripotent stem cells）中过表达（overexpressed）两种小鼠来源的Neurogenin转录因子（Neurogenin transcription factors），仅用4天便获得了具有双极形态（bipolar morphology）的神经元，其纯度超过90%。该高纯度样本使得我们能够对神经发生（neurogenesis）过程中的信使RNA（mRNA）和微小RNA（microRNA）表达谱（expression profiling）进行分析，进而揭示了干细胞向神经元转化过程中涉及的遗传程序（genetic programs）。随后我们对这些表达谱展开分析，以鉴定神经元分化背后的调控网络（regulatory networks）。