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首页> 外文期刊>Journal of Neuroscience Methods >A new approach to neural cell culture for long-term studies.
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A new approach to neural cell culture for long-term studies.

机译：用于长期研究的神经细胞培养的新方法。

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We have developed a new method for culturing cells that maintains their health and sterility for many months. Using conventional techniques, primary neuron cultures seldom survive more than 2 months. Increases in the osmotic strength of media due to evaporation are a large and underappreciated contributor to the gradual decline in the health of these cultures. Because of this and the ever-present likelihood of contamination by airborne pathogens, repeated or extended experiments on any given culture have until now been difficult, if not impossible. We surmounted survival problems by using culture dish lids that form a gas-tight seal, and incorporate a transparent hydrophobic membrane (fluorinated ethylene-propylene) that is selectively permeable to oxygen (O(2)) and carbon dioxide (CO(2)), and relatively impermeable to water vapor. This prevents contamination and greatly reduces evaporation, allowing the use of a non-humidified incubator. We have employed this technique to grow dissociated cortical cultures from rat embryos on multi-electrode arrays. After more than a year in culture, the neurons still exhibit robust spontaneous electrical activity. The combination of sealed culture dishes with extracellular multi-electrode recording and stimulation enables study of development, adaptation, and very long-term plasticity, across months, in cultured neuronal networks. Membrane-sealed dishes will also be useful for the culture of many other cell types susceptible to evaporation and contamination.
机译：我们已经开发出一种新的细胞培养方法，该方法可以维持其健康和无菌许多个月。使用常规技术，原代神经元培养很少能存活超过2个月。由于蒸发而引起的培养基渗透力的增加是造成这些文化健康程度逐渐下降的重要原因，而人们对此并未给予足够的重视。由于这种原因以及空气中病原体污染的可能性一直存在，因此到目前为止，即使不是不可能，对任何给定培养物进行重复或扩展的实验也是困难的。我们通过使用形成气密密封的培养皿盖克服了生存问题，并结合了透明的疏水膜（氟化乙烯-丙烯），该膜可选择性地渗透氧气（O（2））和二氧化碳（CO（2）） ，并且相对不透水蒸气。这样可以防止污染并大大减少蒸发，从而允许使用非加湿的培养箱。我们已经采用了这种技术来从大鼠胚胎在多电极阵列上生长解离的皮质文化。经过一年多的培养，神经元仍然表现出强大的自发电活动。密封培养皿与细胞外多电极记录和刺激相结合，可以在培养的神经元网络中研究跨月的发育，适应和非常长的可塑性。膜密封培养皿也可用于培养许多其他容易蒸发和污染的细胞类型。

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