Formation of three-dimensional tubular endothelial cell networks under defined serum-free cell culture conditions in human collagen hydrogels
Formation of three-dimensional tubular endothelial cell networks under defined serum-free cell culture conditions in human collagen hydrogels
复制标题
DOI：
10.1038/s41598-019-41985-6
复制
发表时间：
2019-04-01
影响因子：
4.6
通讯作者：
Hilfiker, Andres
中科院分区：
文献类型：
--
作者：
Andree, Birgit;Ichanti, Houda;Hilfiker, Andres
管状内皮细胞网络的实现是具有临床相关尺寸的结构的3D组织工程的先决条件，因为细胞的营养受到扩散极限的挑战。3D网络的体外生成通常在使用含血清的细胞培养基和/或动物来源的基质的条件下实现。在这里，通过使用人脐静脉内皮细胞（HUVEC）与人脂肪组织来源的基质细胞（hASC）组合，使用人胶原蛋白I作为水凝胶和脱细胞的猪小肠粘膜下层作为起始基质来产生3D内皮细胞网络。基质胶/大鼠尾胶原I水凝胶用作对照。将所得构建体在无血清培养基或作为对照的内皮生长培养基-2中培养。对内皮细胞网络进行定量，测试管腔形成以及HUVEC和hASC的相互作用。在无血清条件下，与Matrigel/大鼠尾胶原I构建体相比，含有人胶原I的构建体中的管直径略大。所有其他网络参数基本相似。因此，证明了在无血清培养条件下在作为水凝胶的人胶原I中产生3D内皮细胞网络的可行性。总之，所呈现的成就为临床适用结构的生成铺平了道路。
Implementation of tubular endothelial cell networks is a prerequisite for 3D tissue engineering of constructs with clinically relevant size as nourishment of cells is challenged by the diffusion limit. In vitro generation of 3D networks is often achieved under conditions using serum containing cell culture medium and/or animal derived matrices. Here, 3D endothelial cell networks were generated by using human umbilical vein endothelial cells (HUVECs) in combination with human adipose tissue derived stromal cells (hASCs) employing human collagen I as hydrogel and decellularized porcine small intestinal submucosa as starter matrix. Matrigel/rat tail collagen I hydrogel was used as control. Resulting constructs were cultivated either in serum-free medium or in endothelial growth medium-2 serving as control. Endothelial cell networks were quantified, tested for lumen formation, and interaction of HUVECs and hASCs. Tube diameter was slightly larger in constructs containing human collagen I compared to Matrigel/rat tail collagen I constructs under serum-free conditions. All other network parameters were mostly similar. Thereby, the feasibility of generating 3D endothelial cell networks under serum-free culture conditions in human collagen I as hydrogel was demonstrated. In summary, the presented achievements pave the way for the generation of clinical applicable constructs.