Tissue engineering of functional trileaflet heart valves from human marrow stromal cells
Tissue engineering of functional trileaflet heart valves from human marrow stromal cells
复制标题
DOI：
10.1161/01.cir.0000032872.55215.05
复制
发表时间：
2002-09-24
期刊：
影响因子：
37.8
通讯作者：
Turina, MI
中科院分区：
文献类型：
--
作者：
Hoerstrup, SP;Kadner, A;Turina, MI
背景-我们先前展示了成功的组织工程学和基于血管来源细胞的功能性自体心脏瓣膜的植入。人骨髓基质细胞(MSC)具有向多种细胞分化的潜能，临床上易于获得。目的探讨利用骨髓间充质干细胞构建组织工程心脏瓣膜(TEHV)作为替代细胞来源的可行性，以及体外仿生环境对组织分化的影响。方法与结果：分离、扩增人骨髓间充质干细胞，并用流式细胞仪和免疫组织化学方法进行鉴定。将可快速生物吸收聚合物制成的三叶瓣植入MSC，并在脉动流生物反应器中进行体外生长。形态特征包括组织学和电子显微镜(EM)。免疫组织化学分析细胞外基质(ECM)的形成，细胞外基质(ECM)蛋白含量(胶原、糖胺聚糖)和细胞增殖(DNA)。使用Instron(TM)进行生物力学评估。在生物反应器中观察到所有阀门的同步开启和关闭。骨髓间充质干细胞预接种的流式细胞术ASMA、Vimentin阳性，CD31、LDL、CD14阴性。组织学显示TEHV小叶有存活组织和细胞外基质形成。EM显示了活性的、分泌活跃的肌成纤维细胞(肌动蛋白/肌球蛋白细丝、胶原纤维、弹性蛋白)和融合的、均一的组织表面的典型细胞成分。在TEHV小叶中检测到I型、III型、Asma和Vimentin。结论利用体外仿生环境从人骨髓间充质干细胞中分离培养出功能性TEHV是可行的。新生组织具有人自体心脏瓣膜组织的形态特征和力学性能。人骨髓间充质干细胞具有肌成纤维细胞分化的特征。
Background-We previously demonstrated the successful tissue engineering and implantation of functioning autologous heart valves based on vascular-derived cells. Human marrow stromal cells (MSC) exhibit the potential to differentiate into multiple cell-lineages and can be easily obtained clinically. The feasibility of creating tissue engineered heart valves (TEHV) from MSC as an alternative cell source, and the impact of a biomimetic in vitro environment on tissue differentiation was investigated.Methods and Results-Human MSC were isolated, expanded in culture, and characterized by flow-cytometry and immunohistochemistry. Trileaflet heart valves fabricated from rapidly bioabsorbable polymers were seeded with MSC and grown in vitro in a pulsatile-flow-bioreactor. Morphological characterization included histology and electron microscopy (EM). Extracellular matrix (ECM)-formation was analyzed by immunohistochemistry, ECM protein content (collagen, glycosaminoglycan ) and cell proliferation (DNA) were biochemically quantified. Biomechanical evaluation was performed using Instron(TM). In all valves synchronous opening and closing was observed in the bioreactor. Flow-cytometry of MSC pre-seeding was positive for ASMA, vimentin, negative for CD 31, LDL, CD 14. Histology of the TEHV-leaflets demonstrated viable tissue and ECM formation. EM demonstrated cell elements typical of viable, secretionally active myofibroblasts (actin/myosin filaments, collagen fibrils, elastin) and confluent, homogenous tissue surfaces. Collagen types I, III, ASMA, and vimentin were detected in the TEHV-leaflets. Mechanical properties of the TEHV-leaflets were comparable to native tissue.Conclusion-Generation of functional TEHV from human MSC was feasible utilizing a biomimetic in vitro environment. The neo-tissue showed morphological features and mechanical properties of human native-heart-valve tissue. The human MSC demonstrated characteristics of myofibroblast differentiation.