Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction
Modelling physiological and pathological conditions to study pericyte biology in brain function and dysfunction
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模拟生理和病理条件以研究大脑功能和功能障碍的周细胞生物学
作者：
Justin Rustenhoven;Leon C. D. Smyth;D. Jansson;P. Schweder;M. Aalderink;Emma L. Scotter;E. Mee;R. Faull;T. Park;M. Dragunow
脑周细胞包裹内皮细胞，有助于血脑屏障的形成和维持。此外，周细胞参与CNS免疫应答的几个方面，包括瘢痕形成、粘附分子表达、趋化因子分泌和吞噬作用。体外培养通常用于研究脑周细胞的这些功能，然而，这些是高度可塑性的细胞，并且可以根据其培养条件显示不同的表型和功能反应。在这里，我们试图研究两种常用的培养基，含DMEM/F12的高血清和含周细胞培养基（ScienCell）的低血清，如何改变人脑周细胞的表型和神经炎症反应。从成人脑活检组织中分离周细胞，并在DMEM/F12（D-周细胞）或周细胞培养基（P-周细胞）中培养。使用免疫细胞化学、qRT-PCR和EdU掺入来确定这如何改变其基础表型，包括周细胞标志物的表达、增殖和细胞形态。为了确定培养基是否改变了人脑周细胞中的炎症反应，使用免疫细胞化学、qRT-PCR、细胞计数珠阵列和流式细胞术研究转录因子诱导、趋化因子分泌、粘附分子表达、迁移、吞噬作用和对炎症相关生长因子的反应。P-周细胞显示升高的增殖和一个独特的双极形态相比，D-周细胞。此外，P-周细胞显示周细胞相关标志物NG 2、PDGFRβ和纤连蛋白表达较低，αSMA、CD 146、P4 H和结蛋白表达显著降低，Col-IV表达较高。在两种培养物中均观察到响应于IL-1β刺激的核NF-κ B易位，然而，P-周细胞显示转录因子C/EBPδ表达升高，而粘附分子ICAM-1表达降低。P-周细胞表现出较高的吞噬和迁移能力。两种培养物对生长因子TGFβ1和PDGF-BB的刺激反应相似。尽管它们的表型和响应幅度存在差异，但P-周细胞和D-周细胞对所有检查的功能都有类似的响应，表明这些细胞的神经炎性表型对培养条件是稳健的。
Brain pericytes ensheathe the endothelium and contribute to formation and maintenance of the blood–brain-barrier. Additionally, pericytes are involved in several aspects of the CNS immune response including scarring, adhesion molecule expression, chemokine secretion, and phagocytosis. In vitro cultures are routinely used to investigate these functions of brain pericytes, however, these are highly plastic cells and can display differing phenotypes and functional responses depending on their culture conditions. Here we sought to investigate how two commonly used culture media, high serum containing DMEM/F12 and low serum containing Pericyte Medium (ScienCell), altered the phenotype of human brain pericytes and neuroinflammatory responses. Pericytes were isolated from adult human brain biopsy tissue and cultured in DMEM/F12 (D-pericytes) or Pericyte Medium (P-pericytes). Immunocytochemistry, qRT-PCR, and EdU incorporation were used to determine how this altered their basal phenotype, including the expression of pericyte markers, proliferation, and cell morphology. To determine whether culture media altered the inflammatory response in human brain pericytes, immunocytochemistry, qRT-PCR, cytometric bead arrays, and flow cytometry were used to investigate transcription factor induction, chemokine secretion, adhesion molecule expression, migration, phagocytosis, and response to inflammatory-related growth factors. P-pericytes displayed elevated proliferation and a distinct bipolar morphology compared to D-pericytes. Additionally, P-pericytes displayed lower expression of pericyte-associated markers NG2, PDGFRβ, and fibronectin, with notably lower αSMA, CD146, P4H and desmin, and higher Col-IV expression. Nuclear NF-kB translocation in response to IL-1β stimulation was observed in both cultures, however, P-pericytes displayed elevated expression of the transcription factor C/EBPδ, and lower expression of the adhesion molecule ICAM-1. P-pericytes displayed elevated phagocytic and migratory ability. Both cultures responded similarly to stimulation by the growth factors TGFβ1 and PDGF-BB. Despite differences in their phenotype and magnitude of response, both P-pericytes and D-pericytes responded similarly to all examined functions, indicating that the neuroinflammatory phenotype of these cells is robust to culture conditions.
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影响因子：
3.1
作者：
Balabanov, R;Washington, R;DoreDuffy, P
通讯作者：
DoreDuffy, P
影响因子：
4.2
作者：
C. Brighton;D. Lorich;R. Kupcha;T. Reilly;A. R. Jones;R. A. Woodbury
通讯作者：
C. Brighton;D. Lorich;R. Kupcha;T. Reilly;A. R. Jones;R. A. Woodbury
影响因子：
20.3
作者：
Stratman, Amber N.;Malotte, Kristine M.;Davis, George E.
通讯作者：
Davis, George E.
影响因子：
6
作者：
Baluk, P;Morikawa, S;McDonald, DM
通讯作者：
McDonald, DM
影响因子：
3.1
作者：
Tigges U;Welser-Alves JV;Boroujerdi A;Milner R
通讯作者：
Milner R