Optimized Primary Culture of Neuronal Populations for Subcellular Omics Applications.
Optimized Primary Culture of Neuronal Populations for Subcellular Omics Applications.
复制标题
用于亚细胞组学应用的神经元群的优化原代培养。
DOI：
10.1007/978-1-0716-3401-1_7
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Taylor R
中科院分区：
文献类型：
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作者：
Taylor R
原代细胞培养是一种非常宝贵的方法，经常用于克服与体内实验相关的挑战。在斑马鱼研究中，由于胚胎的高光学透明度，体内活体成像实验是常规的，因此，原代细胞培养的利用较少。然而，该方法仍然具有强大的优势，强调了复杂的斑马鱼细胞培养方案的重要性。在这里，我们提出了一个增强的协议，为24 hpf斑马鱼胚胎的解离产生的原代细胞培养。我们包括一种新的细胞培养基配方，特别有利于神经元的生长和存活，使相对长期的文化。我们概述了玻璃盖玻片上的原代斑马鱼神经元培养，以及在transwell插入物中分离神经突组织用于亚细胞转录组研究等实验。
Primary cell culture is an invaluable method frequently used to overcome challenges associated with in vivo experiments. In zebrafish research, in vivo live imaging experiments are routine owing to the high optical transparency of embryos, and, as a result, primary cell culture has been less utilized. However, the approach still boasts powerful advantages, emphasizing the importance of sophisticated zebrafish cell culture protocols. Here, we present an enhanced protocol for the generation of primary cell cultures by dissociation of 24 hpf zebrafish embryos. We include a novel cell culture medium recipe specifically favoring neuronal growth and survival, enabling relatively long-term culture. We outline primary zebrafish neuronal culture on glass coverslips, as well as in transwell inserts which allow isolation of neurite tissue for experiments such as investigating subcellular transcriptomes.