A novel and simple method for culturing pericytes from mouse brain.
A novel and simple method for culturing pericytes from mouse brain.
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DOI：
10.1016/j.mvr.2012.03.008
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发表时间：
2012-07
影响因子：
3.1
通讯作者：
Milner R
中科院分区：
文献类型：
--
作者：
Tigges U;Welser-Alves JV;Boroujerdi A;Milner R
周细胞在血管的发育、成熟和重塑中起关键作用，并且在中枢神经系统（CNS）中，证据表明周细胞还调节血流并形成血脑屏障的组成部分。由于缺乏周细胞特异性标记物以及难以培养足够数量的高纯度周细胞，这一重要细胞类型的研究受到阻碍。在这里，我们提出了一种新颖而简单的方法来分离和培养大量的周细胞从小鼠中枢神经系统，但导致非常纯的周细胞文化。在我们的方法中，从成年小鼠脑中获得的血管细胞最初在针对内皮细胞优化的条件下培养，但在两次传代后切换到针对周细胞生长优化的培养基。将细胞再培养1-2代后，我们获得了基本上同质的细胞群，其表达周细胞标志物NG 2、血小板衍生生长因子受体和CD 146，但内皮细胞标志物（CD 31）、小胶质细胞（Mac-1）和星形胶质细胞（GFAP）呈阴性。在这些条件下，周细胞可以生长到高传代次数，并保持高度纯，很大程度上未分化，确定抗原表达谱和低水平的-SMA表达，周细胞分化的标志物。此外，将细胞从周细胞培养基切换到含有10%FBS的DMEM中促进-SMA表达，表明高传代周细胞仍然可以分化。因此，我们提供了一种替代的方法来培养CNS周细胞，这是很容易建立，并提供了大量的高纯度周细胞延长的时间。这个系统应该为其他在周细胞领域工作的人提供一个有用的额外工具来研究这种迷人的细胞类型的行为。
Pericytes play critical roles in the development, maturation and remodeling of blood vessels, and in the central nervous system (CNS), evidence suggests that pericytes also regulate blood flow and form an integral part of the blood-brain barrier. The study of this important cell type has been hampered by the lack of any pericyte-specific marker and by the difficulty of culturing pericytes in adequate numbers to high purity. Here we present a novel yet simple approach to isolate and culture large numbers of pericytes from the mouse CNS that nevertheless leads to very pure pericyte cultures. In our method, vascular cells obtained from adult mice brains are cultured initially under conditions optimized for endothelial cells, but after two passages switched to a medium optimized for pericyte growth. After growing the cells for 1-2 additional passages we obtained a largely homogeneous population of cells that expressed the pericyte markers NG2, PDGF -receptor, and CD146, but were negative for markers of endothelial cells (CD31), microglia (Mac-1) and astrocytes (GFAP). Under these conditions, pericytes could be grown to high passage number, and were maintained highly pure and largely undifferentiated, as determined by antigen expression profile and low levels of -SMA expression, a marker of pericyte differentiation. Furthermore, switching the cells from pericyte medium into DMEM containing 10% FBS promoted -SMA expression, demonstrating that high passage pericytes could still differentiate. Thus, we provide an alternative approach to the culture of CNS pericytes that is easy to establish and provides large numbers of highly pure pericytes for extended periods of time. This system should provide others working in the pericyte field with a useful additional tool to study the behavior of this fascinating cell type.
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影响因子：
37.8
作者：
Farrington-Rock, C;Crofts, NJ;Canfield, AE
通讯作者：
Canfield, AE
影响因子：
2.6
作者：
Covas, Dimas T.;Panepucci, Rodrigo A.;Zago, Marco A.
通讯作者：
Zago, Marco A.
DOI：
10.1023/a:1011965307612
发表时间：
2001-01-01
期刊：
JOURNAL OF NEUROCYTOLOGY
影响因子：
--
作者：
Bandopadhyay, R;Orte, C;Allt, G
通讯作者：
Allt, G
影响因子：
2.6
作者：
Andreeva, ER;Pugach, IM;Orekhov, AN
通讯作者：
Orekhov, AN
影响因子：
21.3
作者：
Dellavalle, Arianna;Sampaolesi, Maurilio;Cossu, Giulio
通讯作者：
Cossu, Giulio