Towards neuronal organoids: a method for long-term culturing of high-density hippocampal neurons.
Towards neuronal organoids: a method for long-term culturing of high-density hippocampal neurons.
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DOI：
10.1371/journal.pone.0058996
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Patterson RL
中科院分区：
文献类型：
--
作者：
Todd GK;Boosalis CA;Burzycki AA;Steinman MQ;Hester LD;Shuster PW;Patterson RL
神经科学的目标之一是获得易于处理的实验室培养物，在保持实验室易用性的同时，紧密地概括体内系统。由于神经元可以在体内终生存在，因此需要长期的培养系统，以接近实验室培养条件下的生理条件。理想情况下，这样的神经元器官培养应该包含多种细胞类型，高度分化，并具有高密度的相互连接的细胞。然而，在创造这些类型的培养之前，必须解决与长期神经元培养相关的某些问题。我们试图开发一种新的方案，可以进一步延长E18大鼠海马区培养的持续时间和完整性。我们已经开发出一种方案，允许高密度培养E18海马神经元超过120天。这些培养的海马神经元(I)分化良好，具有大量突触，(Ii)牢固地固定在它们的底物上，(Iii)具有高水平的功能连接，(Iv)形成致密的多层细胞网络。我们认为，我们的培养方法可能对多种神经元亚型有效-特别是那些可以在Neurobasal/B27培养液中生长的神经元亚型。这种方法为神经元的长期功能研究提供了新的途径。
One of the goals in neuroscience is to obtain tractable laboratory cultures that closely recapitulate in vivo systems while still providing ease of use in the lab. Because neurons can exist in the body over a lifetime, long-term culture systems are necessary so as to closely mimic the physiological conditions under laboratory culture conditions. Ideally, such a neuronal organoid culture would contain multiple cell types, be highly differentiated, and have a high density of interconnected cells. However, before these types of cultures can be created, certain problems associated with long-term neuronal culturing must be addressed. We sought to develop a new protocol which may further prolong the duration and integrity of E18 rat hippocampal cultures. We have developed a protocol that allows for culturing of E18 hippocampal neurons at high densities for more than 120 days. These cultured hippocampal neurons are (i) well differentiated with high numbers of synapses, (ii) anchored securely to their substrate, (iii) have high levels of functional connectivity, and (iv) form dense multi-layered cellular networks. We propose that our culture methodology is likely to be effective for multiple neuronal subtypes–particularly those that can be grown in Neurobasal/B27 media. This methodology presents new avenues for long-term functional studies in neurons.
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