ABSTRACT

Vascularisation is essential for the development of tailored, tissue-engineered organs and tissues due to diffusion limits of nutrients and the lack of the necessary connection to the cardiovascular system. To pre-vascularize, endothelial cells and supporting cells can be embedded in the scaffold to foster an adequate nutrient and oxygen supply after transplantation. This technique is applied for tissue engineering of various tissues, but there have been few studies on the use of different cell types or cells sources. We compare the effect of supporting cells from different sources on vascularisation. Fibrin gels and agarose-collagen hydrogels were used as scaffolds. The supporting cells were primary human dermal fibroblasts (HDFs), human nasal fibroblasts (HNFs), human mesenchymal stem cells from umbilical cord’s Wharton’s jelly (WJ MSCs), adipose-derived MSCs (AD MSCs) and femoral bone marrow-derived MSCs (BM MSCs). The tissue constructs were incubated for 14 days and analyzed by two-photon laser scanning microscopy. Vascularisation was supported by all cell types, forming branched networks of tubular vascular structures in both hydrogels. In general, fibrin gels present a higher angiogenic promoting environment compared to agarose-collagen hydrogels and fibroblasts show a high angiogenic potential in co-culture with endothelial cells. In agarose-collagen hydrogels, vascular structures supported by AD MSCs were comparable to our HDF control in terms of volume, area and length. BM MSCs formed a homogeneous network of smaller structures in both hydrogels. This study provides data toward understanding the pre-vascularisation properties of different supporting cell types and sources for tissue engineering of different organs and tissues.

摘要

由于营养素的扩散限制以及与心血管系统缺乏必要的联系，血管化对于定制的组织工程器官和组织的发育至关重要。为了预血管化，内皮细胞和支持细胞可以嵌入支架中，以在移植后培养足够的营养和氧气供应。该技术应用于各种组织的组织工程，但对不同细胞类型或细胞来源的使用研究很少。我们比较了来自不同来源的支持细胞对血管形成的影响。纤维蛋白凝胶和琼脂糖-胶原水凝胶用作支架。支持细胞是原代人真皮成纤维细胞 (HDF)、人鼻成纤维细胞 (HNF)、来自脐带沃顿氏胶 (WJ MSC) 的人间充质干细胞、脂肪来源的 MSC (AD MSC) 和股骨骨髓来源的 MSC (BM MSC)。将组织构建体孵育 14 天并通过双光子激光扫描显微镜进行分析。所有细胞类型都支持血管形成，在两种水凝胶中形成管状血管结构的分支网络。一般来说，与琼脂糖-胶原蛋白水凝胶相比，纤维蛋白凝胶具有更高的血管生成促进环境，成纤维细胞在与内皮细胞共培养时显示出高血管生成潜力。在琼脂糖胶原水凝胶中，AD MSCs 支持的血管结构在体积、面积和长度方面与我们的 HDF 对照相当。BM MSCs 在两种水凝胶中形成了较小结构的均匀网络。