Osaki Tatsuya, Serrano Jean C, Kamm Roger D
Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Regen Eng Transl Med. 2018 Sep;4(3):120-132. doi: 10.1007/s40883-018-0054-2. Epub 2018 Apr 23.
In this study, we modeled lymphangiogenesis and vascular angiogenesis in a microdevice using a tissue engineering approach. Lymphatic vessels (LV) and blood vessels (BV) were fabricated by sacrificial molding with seeding human lymphatic endothelial cells and human umbilical vein endothelial cells into molded microchannels (600 μm diameter). During subsequent perfusion culture, lymphangiogenesis and vascular angiogenesis were induced by addition of phorbol 12-myristate 13-acetate (PMA) and VEGF-C or VEGF-A characterized by podoplanin and Prox-1 expression. The lymphatic capillaries formed button-like junctions treated with dexamethasone. To test the potential for screening anti-angiogenic (vascular and lymphatic) factors, antagonists of VEGF were introduced. We found that an inhibitor of VEGF-R3 did not completely suppress lymphatic angiogenesis with BVs present, although lymphatic angiogenesis was selectively prevented by addition of a VEGF-R3 inhibitor without BVs. To probe the mechanism of action, we focus on matrix metalloproteinase (MMP) secretion by vascular endothelial cells and lymphatic endothelial cells under monoculture or co-culture conditions. We found that vascular angiogenesis facilitated lymphangiogenesis via remodeling of the local microenvironment by the increased secretion of MMP, mainly by endothelial cells. Applications of this model include a drug screening assay for corneal disease and models for tumorigenesis including lymphatic angiogenesis and vascular angiogenesis.
在本研究中，我们采用组织工程方法在微器件中模拟淋巴管生成和血管生成。通过牺牲成型法制造淋巴管（LV）和血管（BV），即将人淋巴管内皮细胞和人脐静脉内皮细胞接种到成型的微通道（直径600μm）中。在随后的灌注培养过程中，通过添加佛波醇12 -肉豆蔻酸酯13 -乙酸酯（PMA）和以血小板内皮细胞黏附分子（podoplanin）和Prox - 1表达为特征的VEGF - C或VEGF - A来诱导淋巴管生成和血管生成。经地塞米松处理后，淋巴管毛细血管形成纽扣状连接。为了测试筛选抗血管生成（血管和淋巴管）因子的潜力，引入了VEGF拮抗剂。我们发现，VEGF - R3抑制剂在存在BVs的情况下并未完全抑制淋巴管生成，尽管在没有BVs的情况下添加VEGF - R3抑制剂可选择性地阻止淋巴管生成。为了探究作用机制，我们聚焦于在单培养或共培养条件下血管内皮细胞和淋巴管内皮细胞分泌基质金属蛋白酶（MMP）的情况。我们发现，血管生成通过主要由内皮细胞增加分泌MMP来重塑局部微环境，从而促进淋巴管生成。该模型的应用包括用于角膜疾病的药物筛选试验以及用于肿瘤发生的模型，包括淋巴管生成和血管生成。
