Evaluation of the stromal vascular fraction of adipose tissue as the basis for a stem cell-based tissue-engineered vascular graft.
Evaluation of the stromal vascular fraction of adipose tissue as the basis for a stem cell-based tissue-engineered vascular graft.
复制标题
DOI：
10.1016/j.jvs.2016.09.034
复制
发表时间：
2017-09
影响因子：
4.3
通讯作者：
Vorp DA
中科院分区：
文献类型：
--
作者：
Krawiec JT;Liao HT;Kwan LL;D'Amore A;Weinbaum JS;Rubin JP;Wagner WR;Vorp DA
血管组织工程领域内的限速障碍之一是与在培养物中扩增适当细胞类型相关的冗长的制造时间。用于此目的的一种特别有吸引力的细胞类型是脂肪来源的间充质干细胞（AD-MSC），其丰富且易于从吸脂手术中收获。然而，即使这种细胞类型也有其缺点，包括扩增所需的培养期，这可能造成细胞转化或污染的风险。完全消除文化将是避免这些担忧的理想选择。在这项研究中，我们利用消化人吸脂抽吸物后获得的原始细胞群-称为基质血管组分（SVF）-作为组织工程血管移植物（TEVG）的丰富的无培养细胞来源。首先评估SVF细胞和供体配对培养的AD-MSC在血管紧张素II刺激后分化为血管平滑肌细胞（SMC）和分泌促进SMC迁移的因子（例如条件培养基）的能力。接下来，将两种细胞类型掺入TEVG支架中，作为主动脉移植物植入刘易斯大鼠模型中，并评估其通畅性和组成。通常，来自人SVF的细胞能够执行与通过培养扩增从相同供体分离的AD-MSC相同的功能。具体而言，SVF内的细胞执行两个重要功能，即，它们能够分化成SMC（SVF钙调蛋白表达：16.4% ± 7.7 vs. AD-MSC：19.9% ± 1.7）并且能够分泌促迁移因子（相对于对照的SVF迁移率：3.1 ± 0.3 vs. AD-MSC：2.5 ± 0.5）。此外，SVF还能够被接种在生物可降解的、弹性的、多孔的支架内，当在体内植入8周时，产生了由原代血管组分（例如SMC、内皮细胞、胶原蛋白和弹性蛋白）填充的通畅的TEVG（SVF：83%通畅率对AD-MSC：100%通畅率）。人类脂肪组织可用作无培养细胞来源以产生TEVG，从而为细胞接种移植物的快速生产奠定基础。
One of the rate-limiting barriers within the field of vascular tissue engineering is the lengthy fabrication time associated with expanding appropriate cell types in culture. One particularly attractive cell type for this purpose is the adipose-derived mesenchymal stem cell (AD-MSC), which is abundant and easily harvested from liposuction procedures. However, even this cell type has its drawbacks including the required culture period for expansion which could pose risks of cellular transformation or contamination. Eliminating culture entirely would be ideal to avoid these concerns. In this study we utilized the raw population of cells obtained after digestion of human liposuction aspirates – known as the stromal vascular fraction (SVF) – as an abundant, culture-free cell source for tissue engineered vascular grafts (TEVG). SVF cells and donor-paired cultured AD-MSCs were first assessed for their abilities to differentiate into vascular smooth muscle cells (SMCs) after angiotensin II stimulation and to secrete factors (e.g. conditioned media) that promote SMC migration. Next, both cell types were incorporated into TEVG scaffolds, implanted as an aortic graft in a Lewis rat model, and assessed for their patency and composition. In general, cells from human SVF were able to perform the same functions as AD-MSCs isolated from the same donor via culture expansion. Specifically, cells within the SVF performed two important functions, namely, they were able to differentiate into SMCs (SVF calponin expression: 16.4% ± 7.7 vs. AD-MSC: 19.9% ± 1.7) and could secrete pro-migratory factors (SVF migration rate relative to control: 3.1 ± 0.3 vs. AD-MSC: 2.5 ± 0.5). Additionally, SVF was also capable of being seeded within biodegradable, elastomeric, porous scaffolds that, when implanted in vivo for 8 weeks, generated patent TEVGs (SVF: 83% patency vs. AD-MSC: 100% patency) populated with primary vascular components (e.g. SMCs, endothelial cells, collagen, and elastin). Human adipose tissue can be utilized as a culture-free cell source to create TEVGs, laying the groundwork for the rapid production of cell-seeded grafts.