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首页> 外文期刊>American Journal of Physiology >Potential of fibroblasts to regulate the formation of three-dimensional vessel-like structures from endothelial cells in vitro.
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Potential of fibroblasts to regulate the formation of three-dimensional vessel-like structures from endothelial cells in vitro.

机译：成纤维细胞在体外调节内皮细胞三维血管样结构形成的潜力。

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摘要

The development of vessel-like structures in vitro to mimic as well as to realize the possibility of tissue-engineered small vascular networks presents a major challenge to cell biologists and biotechnologists. We aimed to establish a three-dimensional (3-D) culture system with an endothelial network that does not require artificial substrates or ECM compounds. By using human skin fibroblasts and endothelial cells (ECs) from the human umbilical vein (HUVECs) in diverse spheroid coculture strategies, we verified that fibroblast support and modulate EC migration, viability, and network formation in a 3-D tissue-like stromal environment. In mixed spheroid cultures consisting of human ECs and fibroblasts, a complex 3-D network with EC tubular structures, lumen formation, pinocytotic activity, and tight junction complexes has been identified on the basis of immunohistochemical and transmission electron microscopic imaging. Tubular networks with extensions up to 400 mum were achieved. When EC suspensions were used, EC migration and network formation were critically affected by the status of the fibroblast. However, the absence of EC migration into the center of 14-day, but not 3-day, precultured fibroblast spheroids could not be attributed to loss of F viability. In parallel, it was also confirmed that migrated ECs that entered cluster-like formations became apoptotic, whereas the majority of those forming vessel-like structures remained viable for >8 days. Our protocols allow us to study the nature of tubule formation in a manner more closely related to the in vivo situation as well as to understand the basis for the integration of capillary networks in tissue grafts and develop methods of quantifying the amount of angiogenesis in spheroids using fibroblast and other cells isolated from the same patient, along with ECs.
机译：体外模拟和实现组织工程化小血管网络可能性的血管样结构的发展，对细胞生物学家和生物技术人员提出了重大挑战。我们旨在建立一个不需要人工底物或ECM化合物的具有内皮网络的三维（3-D）培养系统。通过在各种球状共培养策略中使用人皮肤成纤维细胞和人脐静脉（HUVEC）的内皮细胞（EC），我们验证了成纤维细胞在3-D组织样基质环境中支持和调节EC迁移，生存能力和网络形成。在由人类EC和成纤维细胞组成的混合球体培养物中，基于免疫组织化学和透射电子显微镜成像，已鉴定出具有EC管状结构，管腔形成，胞吞活性和紧密连接复合物的复杂3-D网络。实现了扩展到400毫米的管状网络。当使用EC悬浮液时，EC迁移和网络形成受到成纤维细胞状态的严重影响。然而，没有EC迁移到14天而不是3天的中心，预培养的成纤维细胞球体不能归因于F生存力的丧失。同时，还证实进入簇状形成的迁移的ECs凋亡，而大多数形成血管样结构的ECs存活> 8天。我们的协议使我们能够以与体内情况更紧密相关的方式研究肾小管形成的性质，并了解毛细血管网络在组织移植物中的整合基础，并开发出定量方法来使用从同一患者中分离出的成纤维细胞和其他细胞以及EC。

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