工程化人造血管：源自人脐带祖细胞的功能性内皮细胞
Engineered living blood vessels: functional endothelia generated from human umbilical cord-derived progenitors.
作者信息
Schmidt Dörthe, Asmis Lars M, Odermatt Bernhard, Kelm Jens, Breymann Christian, Gössi Matthias, Genoni Michele, Zund Gregor, Hoerstrup Simon P
机构信息
Clinic for Cardiovascular Surgery, University Hospital Zurich, Switzerland.
出版信息
Ann Thorac Surg. 2006 Oct;82(4):1465-71; discussion 1471. doi: 10.1016/j.athoracsur.2006.05.066.
BACKGROUND
Tissue-engineered living blood vessels (TEBV) with growth capacity represent a promising new option for the repair of congenital malformations. We investigate the functionality of TEBV with endothelia generated from human umbilical cord blood-derived endothelial progenitor cells.
METHODS
Tissue-engineered living blood vessels were generated from human umbilical cord-derived myofibroblasts seeded on biodegradable vascular scaffolds, followed by endothelialization with differentiated cord blood-derived endothelial progenitor cells. During in vitro maturation the TEBV were exposed to physiologic conditioning in a flow bioreactor. For functional assessment, a subgroup of TEBV was stimulated with tumor necrosis factor-alpha. Control vessels endothelialized with standard vascular endothelial cells were treated in parallel. Analysis of the TEBV included histology, immunohistochemistry, biochemistry (extracellular matrix analysis, DNA), and biomechanical testing. Endothelia were analyzed by flow cytometry and immunohistochemistry (CD31, von Willebrand factor, thrombomodulin, tissue factor, endothelial nitric oxide synthase).
RESULTS
Histologically, a three-layered tissue organization of the TEBV analogous to native vessels was observed, and biochemistry revealed the major matrix constituents (collagen, proteoglycans) of blood vessels. Biomechanical properties (Young's modulus, 2.03 +/- 0.65 MPa) showed profiles resembling those of native tissue. Endothelial progenitor cells expressed typical endothelial cell markers CD31, von Willebrand factor, and endothelial nitric oxide synthase comparable to standard vascular endothelial cells. Stimulation with tumor necrosis factor-alpha resulted in physiologic upregulation of tissue factor and downregulation of thrombomodulin expression.
CONCLUSIONS
These results indicate that TEBV with tissue architecture and functional endothelia similar to native blood vessels can be successfully generated from human umbilical cord progenitor cells. Thus, blood-derived progenitor cells obtained before or at birth may enable the clinical realization of tissue engineering constructs for pediatric applications.
背景
具有生长能力的组织工程化活性血管（TEBV）是修复先天性畸形的一种有前景的新选择。我们研究了由人脐带血来源的内皮祖细胞生成的带有内皮的TEBV的功能。
方法
将人脐带来源的肌成纤维细胞接种在可生物降解的血管支架上生成组织工程化活性血管，随后用分化的脐带血来源的内皮祖细胞进行内皮化。在体外成熟过程中，TEBV在流动生物反应器中接受生理条件处理。为进行功能评估，用肿瘤坏死因子-α刺激TEBV的一个亚组。同时对用标准血管内皮细胞进行内皮化的对照血管进行平行处理。对TEBV的分析包括组织学、免疫组织化学、生物化学（细胞外基质分析、DNA）和生物力学测试。通过流式细胞术和免疫组织化学（CD31、血管性血友病因子、血栓调节蛋白、组织因子、内皮型一氧化氮合酶）分析内皮。
结果
组织学上，观察到TEBV具有类似于天然血管的三层组织结构，生物化学分析揭示了血管的主要基质成分（胶原蛋白、蛋白聚糖）。生物力学特性（杨氏模量，2.03±0.65兆帕）显示出与天然组织相似的特征。内皮祖细胞表达典型的内皮细胞标志物CD31、血管性血友病因子和内皮型一氧化氮合酶，与标准血管内皮细胞相当。用肿瘤坏死因子-α刺激导致组织因子的生理性上调和血栓调节蛋白表达的下调。
结论
这些结果表明，可以成功地从人脐带祖细胞生成具有类似于天然血管的组织结构和功能性内皮的TEBV。因此，出生前或出生时获得的血源性祖细胞可能使儿科应用的组织工程构建体在临床上得以实现。
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