Chan Xin Yi, Black Rebecca, Dickerman Kayla, Federico Joseph, Lévesque Mathieu, Mumm Jeff, Gerecht Sharon
From the Department of Chemical and Biomolecular Engineering, Institute for NanoBioTechnology (X.Y.C., R.B., K.D., J.F., S.G.) and Department of Materials Science and Engineering (S.G.), Johns Hopkins University, Baltimore, MD; and Department of Ophthalmology, Wilmer Eye Institute (M.L., J.M.) and McKusick-Nathans Institute of Genetic Medicine (M.L., J.M.), Johns Hopkins University School of Medicine, Baltimore, MD.
Arterioscler Thromb Vasc Biol. 2015 Dec;35(12):2677-85. doi: 10.1161/ATVBAHA.115.306362. Epub 2015 Oct 8.
In diabetics, hyperglycemia results in deficient endothelial progenitors and cells, leading to cardiovascular complications. We aim to engineer 3-dimensional (3D) vascular networks in synthetic hydrogels from type 1 diabetes mellitus (T1D) patient-derived human-induced pluripotent stem cells (hiPSCs), to serve as a transformative autologous vascular therapy for diabetic patients.
We validated and optimized an adherent, feeder-free differentiation procedure to derive early vascular cells (EVCs) with high portions of vascular endothelial cadherin-positive cells from hiPSCs. We demonstrate similar differentiation efficiency from hiPSCs derived from healthy donor and patients with T1D. T1D-hiPSC-derived vascular endothelial cadherin-positive cells can mature to functional endothelial cells-expressing mature markers: von Willebrand factor and endothelial nitric oxide synthase are capable of lectin binding and acetylated low-density lipoprotein uptake, form cords in Matrigel and respond to tumor necrosis factor-α. When embedded in engineered hyaluronic acid hydrogels, T1D-EVCs undergo morphogenesis and assemble into 3D networks. When encapsulated in a novel hypoxia-inducible hydrogel, T1D-EVCs respond to low oxygen and form 3D networks. As xenografts, T1D-EVCs incorporate into developing zebrafish vasculature.
Using our robust protocol, we can direct efficient differentiation of T1D-hiPSC to EVCs. Early endothelial cells derived from T1D-hiPSC are functional when mature. T1D-EVCs self-assembled into 3D networks when embedded in hyaluronic acid and hypoxia-inducible hydrogels. The capability of T1D-EVCs to assemble into 3D networks in engineered matrices and to respond to a hypoxic microenvironment is a significant advancement for autologous vascular therapy in diabetic patients and has broad importance for tissue engineering.
在糖尿病患者中，高血糖会导致内皮祖细胞和细胞功能缺陷，进而引发心血管并发症。我们旨在利用1型糖尿病（T1D）患者来源的人诱导多能干细胞（hiPSC）在合成水凝胶中构建三维（3D）血管网络，作为糖尿病患者变革性的自体血管治疗方法。
我们验证并优化了一种无饲养层的贴壁分化程序，以从hiPSC中获得高比例血管内皮钙黏蛋白阳性细胞的早期血管细胞（EVC）。我们证明了来自健康供体和T1D患者的hiPSC具有相似的分化效率。T1D-hiPSC来源的血管内皮钙黏蛋白阳性细胞可成熟为表达成熟标志物的功能性内皮细胞：血管性血友病因子和内皮型一氧化氮合酶能够进行凝集素结合和乙酰化低密度脂蛋白摄取，在基质胶中形成条索并对肿瘤坏死因子-α作出反应。当嵌入工程化透明质酸水凝胶中时，T1D-EVC会发生形态发生并组装成3D网络。当封装在新型缺氧诱导水凝胶中时，T1D-EVC对低氧作出反应并形成3D网络。作为异种移植物，T1D-EVC可整合到发育中的斑马鱼脉管系统中。
使用我们可靠的方案，我们可以将T1D-hiPSC高效分化为EVC。源自T1D-hiPSC的早期内皮细胞成熟时具有功能。当嵌入透明质酸和缺氧诱导水凝胶中时，T1D-EVC可自组装成3D网络。T1D-EVC在工程化基质中组装成3D网络并对缺氧微环境作出反应的能力是糖尿病患者自体血管治疗的一项重大进展，对组织工程具有广泛的重要性。
