Koppensteiner Peter, Boehm Stefan, Arancio Ottavio
1 Department of Pathology & Cell Biology, Columbia University , New York, NY, 10032.
Cell Reprogram. 2014 Dec;16(6):439-46. doi: 10.1089/cell.2014.0054.
The direct conversion of human fibroblasts to neuronal cells, termed human induced neuronal (hiN) cells, has great potential for future clinical advances. However, previous studies have not provided an in-depth analysis of electrophysiological properties of adult fibroblast-derived hiN cultures. We have examined the electrophysiological profile of hiN cells by measuring passive and active membrane properties, as well as spontaneous and evoked neurotransmission. We found that hiN cells exhibited passive membrane properties equivalent to perinatal rodent neurons. In addition, 30% of hiN cells were incapable of action potential (AP) generation and did not exhibit rectifying membrane currents, and none of the cells displayed firing patterns of typical glutamatergic pyramidal neurons. Finally, hiN cells exhibited neither spontaneous nor evoked neurotransmission. Our results suggest that current methods used to produce hiN cells provide preparations in which cells do not achieve the cellular properties of fully mature neurons, rendering these cells inadequate to investigate pathophysiological mechanisms.
将人类成纤维细胞直接转化为神经元细胞，即所谓的人类诱导神经元（hiN）细胞，在未来临床进展方面具有巨大潜力。然而，以往的研究尚未对成体成纤维细胞来源的hiN培养物的电生理特性进行深入分析。我们通过测量被动和主动膜特性以及自发和诱发的神经传递来研究hiN细胞的电生理特征。我们发现hiN细胞表现出与围产期啮齿动物神经元相当的被动膜特性。此外，30%的hiN细胞无法产生动作电位（AP），也未表现出整流膜电流，且没有细胞呈现典型谷氨酸能锥体神经元的放电模式。最后，hiN细胞既不表现出自发神经传递也不表现出诱发神经传递。我们的结果表明，目前用于产生hiN细胞的方法所提供的细胞制剂中，细胞并未达到完全成熟神经元的细胞特性，使得这些细胞不足以用于研究病理生理机制。
