Bin Feng, Yinglong Liu, Nin Xie, Kai Feng, Laifeng Son, Xiaodong Zhu
Department of Thoracic and Cardiovascular Surgery, The Central Hospital of Nanchong, Nanchong, China.
ASAIO J. 2006 May-Jun;52(3):303-9. doi: 10.1097/01.mat.0000206125.81406.02.
Human mesenchymal stem cells (MSCs) differentiate into multiple cell-lineages and may serve as an alternative source of seed cells for tissue engineering. We investigated whether MSCs could be induced to differentiate into endothelial cells (ECs) and function as seed cells for the in vitro construction of tissue-engineered heart valves (TEHVs). Aortic or pulmonary valve homografts were decellularized with 0.1% sodium dodecylsulphate and used as scaffolds for TEHVs. The MSCs were isolated from human bone marrow by Percoll gradient centrifugation (1.073 g/ml), differentiated into ECs with vascular endothelial growth factor (10 ng/ml), and seeded onto a decellularized scaffold (high-density seeding, >10(5) cells/cm2) and grown in static culture for 14 days. Over 90% of the differentiated cells from MSCs stained positively for von Willebrand factor and Tie-2-related antigen. Additionally, Weibel-Palade corpuscle was observed in the cytoplasm of these cells. Levels of reendothelialization in static culture on days 7, 14, and 20, were 73%, 85%, and 95%, respectively. These results show that MSCs from human bone marrow can differentiate, in vitro, into ECs that can then be used to construct TEHVs. Reendothelialization in static culture can be used to provide the basic material for pulsatile-flow cultivation.
人骨髓间充质干细胞（MSCs）可分化为多种细胞谱系，有望成为组织工程种子细胞的替代来源。我们研究了MSCs是否可被诱导分化为内皮细胞（ECs），并作为种子细胞用于体外构建组织工程心脏瓣膜（TEHVs）。用0.1%十二烷基硫酸钠对主动脉或肺动脉瓣同种异体移植物进行去细胞处理，并用作TEHVs的支架。通过Percoll梯度离心法（1.073 g/ml）从人骨髓中分离出MSCs，用血管内皮生长因子（10 ng/ml）将其分化为ECs，然后接种到去细胞支架上（高密度接种，>10⁵个细胞/cm²），并在静态培养中培养14天。超过90%的MSCs分化细胞对血管性血友病因子和Tie-2相关抗原呈阳性染色。此外，在这些细胞的细胞质中观察到了Weibel-Palade小体。在第7天、14天和20天静态培养中的再内皮化水平分别为73%、85%和95%。这些结果表明，人骨髓来源的MSCs在体外可分化为ECs，进而可用于构建TEHVs。静态培养中的再内皮化可为搏动流培养提供基础材料。
