Hematopoietic stem cell cytokines and fibroblast growth factor-2 stimulate human endothelial cell-pericyte tube co-assembly in 3D fibrin matrices under serum-free defined conditions.
Hematopoietic stem cell cytokines and fibroblast growth factor-2 stimulate human endothelial cell-pericyte tube co-assembly in 3D fibrin matrices under serum-free defined conditions.
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DOI：
10.1371/journal.pone.0085147
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Davis GE
中科院分区：
文献类型：
--
作者：
Smith AO;Bowers SL;Stratman AN;Davis GE
我们描述了一种新的三维纤维蛋白基质模型，使用重组造血干细胞细胞因子在无血清条件下促进人类内皮细胞（EC）管与相关周细胞的组装。单个EC和周细胞随机混合在一起，形成EC管，伴随着周细胞在3-5天的时间内聚集到EC管的管腔表面。造血干细胞细胞因子、干细胞因子、白细胞介素-3、基质衍生因子-1α和Flt-3配体与成纤维细胞生长因子(FGF)-2一起添加到纤维蛋白基质中，共同刺激了这些形态发生过程。相反，当纤维蛋白基质中同时加入VEGF和FGF-2时，在无血清条件下，这种管状形成反应不会发生。我们最近证明，VEGF和FGF-2能够对胶原基质中的造血干细胞细胞因子引发EC管的形态发生反应（即在形态发生试验之前添加一夜），有趣的是，它们也能在3D纤维蛋白基质中引发EC管的形态发生。免疫荧光和透射电镜显示，三维纤维蛋白基质中的ec -周细胞相互作用导致血管基底膜的明显组装。此外，我们发现造血干细胞细胞因子和周细胞以依赖于α5β1整合素的方式刺激纤维蛋白基质中的EC发芽。这种新型的共培养系统，在无血清的条件下，允许在关键的ECM环境（即纤维蛋白基质）中对EC管组装、周细胞募集和成熟事件进行分子分析，该环境调节了出生后生命中的血管生成事件。
We describe a novel 3D fibrin matrix model using recombinant hematopoietic stem cell cytokines under serum-free defined conditions which promotes the assembly of human endothelial cell (EC) tubes with co-associated pericytes. Individual ECs and pericytes are randomly mixed together and EC tubes form that is accompanied by pericyte recruitment to the EC tube abluminal surface over a 3-5 day period. These morphogenic processes are stimulated by a combination of the hematopoietic stem cell cytokines, stem cell factor, interleukin-3, stromal derived factor-1α, and Flt-3 ligand which are added in conjunction with fibroblast growth factor (FGF)-2 into the fibrin matrix. In contrast, this tube morphogenic response does not occur under serum-free defined conditions when VEGF and FGF-2 are added together in the fibrin matrices. We recently demonstrated that VEGF and FGF-2 are able to prime EC tube morphogenic responses (i.e. added overnight prior to the morphogenic assay) to hematopoietic stem cell cytokines in collagen matrices and, interestingly, they also prime EC tube morphogenesis in 3D fibrin matrices. EC-pericyte interactions in 3D fibrin matrices leads to marked vascular basement membrane assembly as demonstrated using immunofluorescence and transmission electron microscopy. Furthermore, we show that hematopoietic stem cell cytokines and pericytes stimulate EC sprouting in fibrin matrices in a manner dependent on the α5β1 integrin. This novel co-culture system, under serum-free defined conditions, allows for a molecular analysis of EC tube assembly, pericyte recruitment and maturation events in a critical ECM environment (i.e. fibrin matrices) that regulates angiogenic events in postnatal life.
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影响因子：
9.8
作者：
Astrof S;Hynes RO
通讯作者：
Hynes RO
影响因子：
10.4
作者：
Hynes, R. O.
通讯作者：
Hynes, R. O.
影响因子：
6
作者：
Bayless, KJ;Salazar, R;Davis, GE
通讯作者：
Davis, GE
DOI：
10.1007/978-1-62703-604-7_2
发表时间：
2013-01-01
期刊：
CELL-CELL INTERACTIONS: METHODS AND PROTOCOLS, 2ND EDITION
影响因子：
--
作者：
Davis, George E.;Kim, Dae Joong;Stratman, Amber N.
通讯作者：
Stratman, Amber N.
DOI：
10.1083/jcb.132.6.1177
发表时间：
1996-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Koolwijk P;van Erck MG;de Vree WJ;Vermeer MA;Weich HA;Hanemaaijer R;van Hinsbergh VW
通讯作者：
van Hinsbergh VW