Survival of hippocampal and cortical neurons in a mixture of MEM+ and B27-supplemented neurobasal medium
Survival of hippocampal and cortical neurons in a mixture of MEM+ and B27-supplemented neurobasal medium
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DOI：
10.1016/s0891-5849(99)00268-3
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发表时间：
2000-03-01
影响因子：
7.4
通讯作者：
Lovell, MA
中科院分区：
文献类型：
--
作者：
Xie, CS;Markesbery, WR;Lovell, MA
用无血清添加B-27的neurobasal（NB）和添加10%胎牛血清的Eagle's最低必需培养基（MEM+）培养大鼠胚胎海马神经元。虽然NE培养基导致增强的细胞存活，但其含有生物抗氧化剂，并且不适合于在培养的神经元中研究自由基损伤和氧化。不含额外抗氧化剂的MEM+已被广泛用于自由基损伤和氧化的研究，尽管它不支持培养中的最佳神经元存活。MEM+中的血清导致增强的细胞存活，但也促进神经胶质细胞增殖。在这项研究中，我们使用了一种新的组合培养基（NM-2），包括NE和MEM+生长原代海马和皮质神经元培养。NM-2在密度为50个细胞/mm 2的解离神经元中提高了78.9%的神经元存活率，在密度为100个细胞/mm 2的解离神经元中提高了83.1%的神经元存活率，同时将胶质细胞增殖降低至2-3%，并完全抑制少突胶质细胞。NM-2使介质中抗氧化剂对神经毒素4-羟基壬烯醛的有效性最小化。它还减少了神经元的聚集，并提供了更均匀的神经元分布。神经元在NM-2中存活4周，而不改变原始培养基。NM-2为研究神经元的自由基损伤和氧化提供了良好的环境。该组合结合了NE和MEM+的最佳组合，其导致高神经元存活率、低神经胶质细胞增殖、降低的抗氧化剂水平，并提供海马和皮质神经元的相对纯的培养物。(C)2000 Elsevier Science Inc.
Serum-free B-27 supplemented neurobasal (NB) and a 10% fetal bovine serum-supplemented Eagle's minimum essential medium (MEM+) are used to culture rat embryonic hippocampal neurons for different purposes. Although NE medium leads to enhanced cell survival, it contains biological antioxidants and is not suitable for the study of free radical damage and oxidation in cultured neurons. MEM+ without additional antioxidants has been used widely in the study of free radical damage and oxidation, although it does not support optimum neuronal survival in culture. Serum in MEM+ leads to enhanced cell survival but also promotes glial cell proliferation. In this study, we used a new combination medium (NM-2) that consists of both NE and MEM+ for growing primary hippocampal and cortical neuronal cultures. NM-2 enhanced neuronal survival 78.9% for dissociated neurons at a density of 50 cells/mm(2) and 83.1% for 100 cells/mm(2), while decreasing glial cell proliferation to 2-3% and completely inhibiting oligodendrocytes. The NM-2 minimized the effectiveness of antioxidants in the medium to the neurotoxin 4-hydroxynonenal. It also decreased neuronal clumping and provided a more even distribution of neurons. Neurons survived for 4 weeks in NM-2 without changing the original medium. NM-2 provides a good environment for studies of free radical damage and oxidation of neurons. The combination incorporates the best of both NE and MEM+ that results in high neuron survival rate, low glial cell proliferation, reduced antioxidant level, and provides relatively pure cultures of hippocampal and cortical neurons. (C) 2000 Elsevier Science Inc.