Efficient Generation of Functionally Active Spinal Cord Neurons from Spermatogonial Stem Cells
Efficient Generation of Functionally Active Spinal Cord Neurons from Spermatogonial Stem Cells
复制标题
DOI：
10.1007/s12035-016-0057-2
复制
发表时间：
2016-08
影响因子：
5.1
通讯作者：
Hao Yang;Cui-cui Liu;Bo Chen;Jing An;Rui Zhang;Qian Zhang;Jingjing Zhao;B. He;D. Hao
中科院分区：
文献类型：
--
作者：
Hao Yang;Cui-cui Liu;Bo Chen;Jing An;Rui Zhang;Qian Zhang;Jingjing Zhao;B. He;D. Hao
神经干细胞（NSCs）被认为是细胞移植治疗多节段脊髓损伤和功能恢复的理想候选细胞。然而，可用于损伤移植的神经干细胞的极端缺点仍然是神经再生医学的一个重大瓶颈。因此，必须建立合适的细胞库作为无问题的替代方案。在这里，我们证明，精原干细胞（SSC）来自大鼠睾丸稳健地产生终末分化，功能成熟的脊髓神经元，通过使用优化的分化方案。在进行3周的体外分化程序后，大多数细胞表现出神经形态学特征，Tuj-1阳性。值得注意的是，获得的细胞中约有60%共表达胆碱乙酰转移酶（CHAT）、乙酰胆碱酯酶（AchE）和降钙素基因相关肽（CGRP）。更重要的是，除了获得神经抗原和生物化学特性，几乎所有的神经元有效地表现出类似于野生型神经元的体外功能，如突触形成，增加神经元钙内流，和电生理学。这是第一份揭示从SSC中一致且可重复地产生大量功能性神经元的报告。总的来说，该系统适用于SSC转分化为神经元细胞的研究，并且可以为脊髓损伤的治疗以及遗传和小分子筛选提供足够的神经元。
Neural stem cells (NSCs) are hitherto regarded as perspective candidates for cell transplantation in clinical therapies for multilevel spinal cord injury and function restoration. However, the extreme drawbacks of NSCs available for injury transplantation still represent a significant bottleneck in neural regeneration medicine. Therefore, it is essential to establish a suitable cell reservoir as an issue-free alternative. Here, we demonstrate that spermatogonial stem cells (SSCs) derived from rat testis robustly give rise to terminally differentiated, functionally mature spinal cord neurons by using an optimized differentiation protocol. After performing a 3-week in vitro differentiation procedure, most cells exhibited neural morphological features and were Tuj-1 positive. Of note, approximately 60 % of the obtained cells coexpressed choline acetyltransferase (CHAT), acetylcholinesterase (AchE), and calcitonin gene-related peptide (CGRP). More importantly, apart from acquisition of neural antigenic and biochemical properties, nearly all neurons efficiently exhibited in vitro functionality similar to wild-type neurons, such as synapse formation, increased neuronal calcium influx, and electrophysiology. This is the first report revealing consistent and reproducible generation of large amounts of functional neurons from SSCs. Collectively, this system is suitable for studies of SSC transdifferentiation into neuronal cells and can provide sufficient neurons for the treatment of spinal cord injury as well as for genetic and small molecule screenings.