Contribution of Rat Endothelial Progenitor Cells on Three-Dimensional Network Formation In Vitro
Contribution of Rat Endothelial Progenitor Cells on Three-Dimensional Network Formation In Vitro
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DOI：
10.1089/ten.tea.2008.0417
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发表时间：
2009-09-01
影响因子：
4.1
通讯作者：
Tanishita, Kazuo
中科院分区：
文献类型：
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作者：
Koga, Masaki;Sudo, Ryo;Tanishita, Kazuo
内皮祖细胞（EPC）移植是缺血性疾病的一种有前途的治疗选择，了解EPC介导血管生成作用的机制对于进一步提高其有效性及其在体外组织工程中的应用至关重要。在这里，我们使用体外三维网络模型研究了 EPC 对网络形成的贡献。将牛微血管内皮细胞（EC）接种在胶原凝胶上以创建三维网络模型。随后，将从大鼠骨髓中分离的 EPC 接种在汇合的内皮细胞 (EC) 单层顶部，以创建 EPC + EC 模型。在该模型中，EPC 促进了 EC 网络的形成。定量分析表明，随着 EPC 的加入，网络的总长度、数量和深度显着增加。随着实验持续时间的增加，EPC 倾向于定位在胶原蛋白凝胶内部形成的网络中，而不是定位在汇合的单层上。此外，EPC 优先分布在汇合单层中的芽位置附近。此外，EPC条件培养基促进网络形成，并且在条件培养基中检测到血管内皮生长因子。总而言之，EPC 通过直接结合现场生长因子分泌来促进网络形成。 EPCs 的血管生成能力为体外重建血管组织提供了可能的细胞来源。
Endothelial progenitor cell (EPC) transplantation is a promising treatment option for ischemic diseases, and understanding the mechanism mediating angiogenic effects of EPCs is essential to further improve its effectiveness as well as their application to in vitro tissue engineering. Here we investigated EPC contribution on network formation using an in vitro three-dimensional network model. Bovine microvascular endothelial cells (ECs) were seeded on collagen gel to create a three-dimensional network model. Subsequently, EPCs isolated from rat bone marrow were seeded on top of a confluent endothelial cell (EC) monolayer to create EPC + EC model. In this model, EPCs promoted EC network formation. Quantitative analyses revealed that the total length, number, and depth of networks were significantly enhanced with the addition of EPCs. EPCs tended to localize in networks formed inside collagen gel rather than on a confluent monolayer with increasing experimental duration. In addition, EPCs preferentially distributed near sprout positions in a confluent monolayer. Furthermore, EPC-conditioned medium promoted network formation, and vascular endothelial growth factor was detected in the conditioned medium. Taken together, EPCs contributed to network formation by direct incorporation with on-site growth factor secretion. The angiogenic ability of EPCs offers a possible cell source to reconstruct vascularized tissues in vitro.