TGFbeta is required for the formation of capillary-like structures in three-dimensional cocultures of 10T1/2 and endothelial cells
TGFbeta is required for the formation of capillary-like structures in three-dimensional cocultures of 10T1/2 and endothelial cells
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DOI：
10.1023/a:1016611824696
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发表时间：
2001-01-01
期刊：
影响因子：
9.8
通讯作者：
D'Amore, P. A.
中科院分区：
文献类型：
--
作者：
Darland, D. C.;D'Amore, P. A.
在涉及内皮细胞（EC）和周细胞/平滑肌细胞（SMC）相互作用的过程中，新血管从头（血管发生）或从预先存在的血管（血管生成）形成。这种相互作用的一个基本组成部分是内皮诱导的周细胞和SMC的募集、增殖和随后的分化。我们先前已经证明TGF β诱导C3 H/10 T1/2（10 T1/2）间充质细胞向SMC/周细胞谱系分化。目前的研究测试的假设，TGF β不仅诱导SMC分化，但稳定的毛细血管样结构在体外血管生成的三维（3D）模型。10 T1/2和EC在MatrigelTM中用于建立共培养物，其形成索状结构，使人联想到体内的新毛细血管。在镀覆后2-3小时内开始形成线，并持续至镀覆后18小时。在较长时间的共培养中，索结构分解并形成聚集体。与SMC/周细胞谱系相关的蛋白质如平滑肌α-肌动蛋白（SMA）和NG 2蛋白聚糖的10 T1/2表达在这些3D共培养物中上调。TGF β特异性中和剂的应用阻断了10 T1/2细胞中的索形成并抑制了SMA和NG 2的表达。我们的结论是，TGF β介导的10 T1/2分化为SMC/周细胞在三维共培养，并与分化的壁细胞的MatrigelTM中的毛细血管样结构的形成所需的协会。
New vessels form de novo (vasculogenesis) or from pre-existing vessels (angiogenesis) in a process that involves the interaction of endothelial cells (EC) and pericytes/smooth muscle cells (SMC). One basic component of this interaction is the endothelial-induced recruitment, proliferation and subsequent differentiation of pericytes and SMC. We have previously demonstrated that TGFbeta induces the differentiation of C3H/10T1/2 (10T1/2) mesenchymal cells toward a SMC/pericyte lineage. The current study tests the hypothesis that TGFbeta not only induces SMC differentiation but stabilizes capillary-like structures in a three-dimensional (3D) model of in vitro angiogenesis. 10T1/2 and EC in MatrigelTM were used to establish cocultures that form cord structures that are reminiscent of new capillaries in vivo. Cord formation is initiated within 2-3 h after plating and continues through 18 h after plating. In longer cocultures the cord structures disassemble and form aggregates. 10T1/2 expression of proteins associated with the SMC/pericyte lineage, such as smooth muscle alpha-actin (SMA) and NG2 proteoglycan, are upregulated in these 3D cocultures. Application of neutralizing reagents specific for TGFbeta blocks cord formation and inhibits expression of SMA and NG2 in the 10T1/2 cells. We conclude that TGFbeta mediates 10T1/2 differentiation to SMC/pericytes in the 3D cocultures and that association with differentiated mural cells is required for formation of capillary-like structures in MatrigelTM.