Living Autologous Heart Valves Engineered From Human Prenatally Harvested Progenitors
Living Autologous Heart Valves Engineered From Human Prenatally Harvested Progenitors
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DOI：
10.1161/circulationaha.105.001040
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发表时间：
2006-07
期刊：
影响因子：
37.8
通讯作者：
D. Schmidt;A. Mol;C. Breymann;J. Achermann;Bernhard Odermatt;M. Gössi;S. Neuenschwander;R. Prêtre;M. Genoni;G. Zund;S. Hoerstrup
中科院分区：
文献类型：
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作者：
D. Schmidt;A. Mol;C. Breymann;J. Achermann;Bernhard Odermatt;M. Gössi;S. Neuenschwander;R. Prêtre;M. Genoni;G. Zund;S. Hoerstrup
背景-心脏瓣膜组织工程是克服自体生长替代物缺乏的一种很有前途的策略，特别是用于先天性畸形的修复。在这里，我们提出了一个新的概念，利用人类产前祖细胞作为新的和唯一的细胞来源，以产生准备在出生时使用的自体植入物。方法和结果-从常规取样的产前绒毛膜绒毛标本中分离出人胎儿间充质祖细胞并在体外扩增。其中一部分冷冻保存。表型分型和基因分型后，将细胞播种到合成的可生物降解的小叶支架上（n=12），并在生物反应器中进行调节。21天后，用脐带血来源的内皮祖细胞对小叶进行内皮化，并再培养7天。通过组织学、免疫组织化学、生物化学（细胞外基质、DNA的数量）、力学测试和扫描电镜（SEM）对所得组织进行分析，并与天然新生儿心脏瓣膜小叶进行比较。新鲜和冷冻保存的细胞表现出类似的肌成纤维细胞样表型。基因分型证实了它们的胎儿起源。新组织的组织结构、细胞表型、细胞外基质的产生和DNA含量与它们的原生组织相当。小叶表面覆盖有功能性内皮。扫描电镜显示聚合物表面呈细胞状分布，表面光滑。力学特征与天然心脏瓣膜相似。结论：从常规绒毛膜绒毛取样中获得的产前胎儿祖细胞成功地用作活体心脏瓣膜小叶工程的独家新细胞来源。这一概念可能使具有生长潜力的自体替代物在出生时就可以使用。结合细胞银行技术的使用，这种方法也可以应用于产后应用。
Background— Heart valve tissue engineering is a promising strategy to overcome the lack of autologous growing replacements, particularly for the repair of congenital malformations. Here, we present a novel concept using human prenatal progenitor cells as new and exclusive cell source to generate autologous implants ready for use at birth. Methods and Results— Human fetal mesenchymal progenitors were isolated from routinely sampled prenatal chorionic villus specimens and expanded in vitro. A portion was cryopreserved. After phenotyping and genotyping, cells were seeded onto synthetic biodegradable leaflet scaffolds (n=12) and conditioned in a bioreactor. After 21 days, leaflets were endothelialized with umbilical cord blood-derived endothelial progenitor cells and conditioned for additional 7 days. Resulting tissues were analyzed by histology, immunohistochemistry, biochemistry (amounts of extracellular matrix, DNA), mechanical testing, and scanning electron microscopy (SEM) and were compared with native neonatal heart valve leaflets. Fresh and cryopreserved cells showed comparable myofibroblast-like phenotypes. Genotyping confirmed their fetal origin. Neo-tissues exhibited organization, cell phenotypes, extracellular matrix production, and DNA content comparable to their native counterparts. Leaflet surfaces were covered with functional endothelia. SEM showed cellular distribution throughout the polymer and smooth surfaces. Mechanical profiles approximated those of native heart valves. Conclusions— Prenatal fetal progenitors obtained from routine chorionic villus sampling were successfully used as an exclusive, new cell source for the engineering of living heart valve leaflets. This concept may enable autologous replacements with growth potential ready for use at birth. Combined with the use of cell banking technology, this approach may be applied also for postnatal applications.