A novel method to establish microglia-free astrocyte cultures: comparison of matrix metalloproteinase expression profiles in pure cultures of astrocytes and microglia.
A novel method to establish microglia-free astrocyte cultures: comparison of matrix metalloproteinase expression profiles in pure cultures of astrocytes and microglia.
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DOI：
10.1002/glia.20689
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发表时间：
2008-08-15
期刊：
影响因子：
6.2
通讯作者：
Milner, Richard
中科院分区：
文献类型：
--
作者：
Crocker, Stephen J.;Frausto, Ricardo F.;Whitton, J. Lindsay;Milner, Richard
基质金属蛋白酶（MMP）蛋白水解活性的增加与中枢神经系统中许多神经炎症和神经退行性疾病的发病机制有关。为了充分了解这一过程，确定特定细胞类型的MMP表达谱是很重要的，包括中枢神经系统驻留细胞星形胶质细胞和小胶质细胞。虽然先前的研究通过使用混合胶质细胞培养物来表征星形胶质细胞MMP表达，但这些结果可能因这些培养物中存在污染的小胶质细胞而复杂化。在目前的研究中，我们试图通过一种新的方法来澄清这种复杂性，通过促进神经干细胞（NSC）向星形胶质细胞的分化，制备完全缺乏小胶质细胞的纯星形胶质细胞培养物。通过RNase保护实验对混合胶质细胞培养物、神经球来源的星形胶质细胞和纯小胶质细胞的MMP表达谱进行了表征。这表明MMP基因的表达在很大程度上是细胞类型特异性的。星形胶质细胞组成性地表达MMP-11、MMP-14和MMP-2，并诱导MMP-3响应IL-1β，但对脂多糖（LPS）无反应。相比之下，小胶质细胞组成性地表达高水平的MMP-12，并表现出对LPS的强烈诱导MMP-9和MMP-14。明胶酶谱证实，LPS和TNF-α诱导MMP-9在小胶质细胞中强表达，而星形胶质细胞中不表达。总之，这些研究表明，神经球来源的星形胶质细胞是体外研究星形胶质细胞行为的一个有吸引力的替代系统。利用这个系统，我们已经证明星形胶质细胞和小胶质细胞表达不同的MMP基因，并且在LPS或TNF-α反应中，小胶质细胞而不是星形胶质细胞是MMP-9的主要来源。
Increased matrix metalloproteinase (MMP) proteolytic activity contributes to the pathogenesis of many neuroinflammatory and neurodegenerative conditions in the CNS. To fully understand this process, it is important to define the MMP expression profile of specific cell types, including the CNS-resident cells astrocytes and microglia. While previous studies have characterized astrocyte MMP expression by using mixed glial cultures, these results are likely complicated by the presence of contaminating microglia within these cultures. In the current study, we sought to clarify this complexity, by taking a novel approach to prepare pure astrocyte cultures entirely devoid of microglia, by promoting neural stem cell (NSC) differentiation into astrocytes. The MMP expression profile of mixed glial cultures, neurosphere-derived astrocytes, and pure microglia was characterized by RNase protection assay. This revealed that MMP gene expression is largely cell-type specific. Astrocytes constitutively expressed MMP-11, MMP-14, and MMP-2 and showed induction of MMP-3 in response to IL-1β but did not respond to lipopolysaccharide (LPS). In contrast, microglia constitutively expressed high levels of MMP-12 and showed strong induction of MMP-9 and MMP-14 in response to LPS. Gelatin zymography confirmed that LPS and TNF-α induced strong expression of MMP-9 in microglia but not astrocytes. In summary, these studies demonstrate that neurosphere-derived astrocytes represent an attractive alternative system in which to study astrocyte behavior in vitro. Using this system, we have shown that astrocytes and microglia express distinct sets of MMP genes and that microglia, not astrocytes, are the major source of MMP-9 in response to LPS or TNF-α.
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DOI：
10.1097/00004647-199906000-00005
发表时间：
1999-06-01
影响因子：
6.3
作者：
Heo, JH;Lucero, J;del Zoppo, GJ
通讯作者：
del Zoppo, GJ
影响因子：
15.9
作者：
Dubois, B;Masure, S;Arnold, B
通讯作者：
Arnold, B
影响因子：
6.2
作者：
Brunet, JF;Grollimund, L;Pellerin, L
通讯作者：
Pellerin, L
影响因子：
5.3
作者：
Kim, YS;Kim, SS;Joh, TH
通讯作者：
Joh, TH
DOI：
10.1111/j.1365-2990.1997.tb01315.x
发表时间：
1997-10-01
影响因子：
5
作者：
Anthony, DC;Ferguson, B;Perry, VH
通讯作者：
Perry, VH