Chen Yucui, Stevens Beth, Chang Jufang, Milbrandt Jeffrey, Barres Ben A, Hell Johannes W
Department of Pharmacology, University of Iowa, Iowa City, IA 52242-1109, United States.
J Neurosci Methods. 2008 Jun 30;171(2):239-47. doi: 10.1016/j.jneumeth.2008.03.013. Epub 2008 Apr 1.
In vitro culturing of primary neurons is a mainstay of neurobiological research. Many of these culture paradigms have taken advantage of defined culture media rather than serum additives that contain undefined survival factors to facilitate experimental manipulations and interpretation of the results. To culture neurons in the absence of serum, defined supplements such as B27 are now widely used. However, commercially available supplements exhibit large variability in their capabilities to support neurons in culture. We re-optimized and modified earlier published formulations of B27 using 21 different ingredients (NS21). NS21 supports neuronal cultures of high quality as manifested by their morphological characteristics, formation of synapses, and postsynaptic responses. Much of the variability in the quality of B27/NS21 was due to variability in the quality of different sources of bovine serum albumin. Furthermore, we found that holo-transferrin used in NS21 is preferable over apo-transferrin used in B27 for the quality of neuronal cultures.
原代神经元的体外培养是神经生物学研究的支柱。许多这些培养范式利用了限定培养基，而非含有未定义存活因子的血清添加剂，以促进实验操作和结果解读。为了在无血清条件下培养神经元，诸如B27等限定补充剂如今被广泛使用。然而，市售补充剂在支持培养神经元的能力方面表现出很大的变异性。我们使用21种不同成分重新优化并修改了早期发表的B27配方（NS21）。NS21支持高质量的神经元培养，这体现在其形态特征、突触形成和突触后反应上。B27/NS21质量的很大一部分变异性是由于不同来源牛血清白蛋白质量的变异性。此外，我们发现NS21中使用的全转铁蛋白比B27中使用的脱铁转铁蛋白更有利于神经元培养的质量。
