Kusuma Sravanti, Facklam Amanda, Gerecht Sharon
1 Department of Chemical and Biomolecular Engineering, Johns Hopkins Physical Sciences-Oncology Center, Institute for NanoBioTechnology, Johns Hopkins University , Baltimore, Maryland.
Stem Cells Dev. 2015 Feb 15;24(4):451-8. doi: 10.1089/scd.2014.0377. Epub 2014 Oct 27.
Tissue-engineered constructs are rendered useless without a functional vasculature owing to a lack of nutrients and oxygen. Cell-based approaches to reconstruct blood vessels can yield structures that mimic native vasculature and aid transplantation. Vascular derivatives of human induced pluripotent stem cells (hiPSCs) offer opportunities to generate patient-specific therapies and potentially provide unlimited amounts of vascular cells. To be used in engineered vascular constructs and confer therapeutic benefit, vascular derivatives must exhibit additional key properties, including extracellular matrix (ECM) production to confer structural integrity and growth factor production to facilitate integration. In this study, we examine the hypothesis that vascular cells derived from hiPSCs exhibit these critical properties to facilitate their use in engineered tissues. hiPSCs were codifferentiated toward early vascular cells (EVCs), a bicellular population of endothelial cells (ECs) and pericytes, under varying low-oxygen differentiation conditions; subsequently, ECs were isolated and passaged. We found that EVCs differentiated under low-oxygen conditions produced copious amounts of collagen IV and fibronectin as well as vascular endothelial growth factor and angiopoietin 2. EVCs differentiated under atmospheric conditions did not demonstrate such abundant ECM expression, but exhibited greater expression of angiopoietin 1. Isolated ECs could proliferate up to three passages while maintaining the EC marker vascular endothelial cadherin. Isolated ECs demonstrated an increased propensity to produce ECM compared with their EVC correlates and took on an arterial-like fate. These findings illustrate that hiPSC vascular derivates hold great potential for therapeutic use and should continue to be a preferred cell source for vascular construction.
由于缺乏营养和氧气,没有功能性脉管系统的组织工程构建体将变得毫无用处。基于细胞的血管重建方法可以产生模仿天然脉管系统并有助于移植的结构。人类诱导多能干细胞(hiPSC)的血管衍生物为产生针对患者的疗法提供了机会,并有可能提供无限数量的血管细胞。为了用于工程化血管构建体并带来治疗益处,血管衍生物必须展现出其他关键特性,包括产生细胞外基质(ECM)以赋予结构完整性,以及产生生长因子以促进整合。在本研究中,我们检验了这样一个假设,即源自hiPSC的血管细胞具有这些关键特性,便于其在工程组织中使用。在不同的低氧分化条件下,hiPSC被共分化为早期血管细胞(EVC),这是一种由内皮细胞(EC)和周细胞组成的双细胞群体;随后,分离并传代EC。我们发现,在低氧条件下分化的EVC产生了大量的IV型胶原蛋白、纤连蛋白以及血管内皮生长因子和血管生成素2。在大气条件下分化的EVC没有表现出如此丰富的ECM表达,但血管生成素1的表达更高。分离的EC可以传代多达三次,同时保持EC标志物血管内皮钙黏蛋白。与它们的EVC对应物相比,分离的EC表现出更强的产生ECM的倾向,并呈现出动脉样命运。这些发现表明,hiPSC血管衍生物在治疗应用方面具有巨大潜力,应继续作为血管构建的首选细胞来源。