In Vitro and In Vivo Preclinical Testing of Pericyte-Engineered Grafts for the Correction of Congenital Heart Defects
In Vitro and In Vivo Preclinical Testing of Pericyte-Engineered Grafts for the Correction of Congenital Heart Defects
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DOI：
10.1161/jaha.119.014214
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发表时间：
2020-02-18
影响因子：
5.4
通讯作者：
Madeddu, Paolo
中科院分区：
文献类型：
--
作者：
Alvino, Valeria Vincenza;Kilcooley, Michael;Madeddu, Paolo
背景我们以前曾报道过的可能性，使用周细胞的姑息手术的遗留物的先天性心脏病工程师临床认证的prosthetic grafts.Methods and Results在这里，我们评估了使用人工管道工程与新生猪周细胞重建肺动脉的9周龄小猪的可行性。在分离和培养扩增后，人和猪心脏周细胞在心脏中的解剖定位和抗原谱方面相似。与人周细胞一样，猪替代物在单细胞分选后形成克隆，分泌血管生成因子和细胞外基质蛋白，并支持体外内皮细胞迁移和网络形成。将接种或未接种（对照）的猪周细胞CorMatrix导管在静态条件下培养5天，然后将其成形为导管并在流动生物反应器中孵育1或2周。免疫组化研究表明，在管道外层的周细胞的活力和整合。机械测试记录了与未接种导管相比，接种导管在最大载荷下的刚度降低和应变增加。然后使用对照和周细胞工程化的导管来替换小猪的左肺动脉。4个月后，解剖和功能整合的移植物被证实使用多普勒超声心动图，心脏磁共振成像，和histologic.Conclusions这些研究结果表明，使用新生儿心脏周细胞重建的小尺寸分支肺动脉在一个大的动物模型的可行性。
Background We have previously reported the possibility of using pericytes from leftovers of palliative surgery of congenital heart disease to engineer clinically certified prosthetic grafts.Methods and Results Here, we assessed the feasibility of using prosthetic conduits engineered with neonatal swine pericytes to reconstruct the pulmonary artery of 9-week-old piglets. Human and swine cardiac pericytes were similar regarding anatomical localization in the heart and antigenic profile following isolation and culture expansion. Like human pericytes, the swine surrogates form clones after single-cell sorting, secrete angiogenic factors, and extracellular matrix proteins and support endothelial cell migration and network formation in vitro. Swine pericytes seeded or unseeded (control) CorMatrix conduits were cultured under static conditions for 5 days, then they were shaped into conduits and incubated in a flow bioreactor for 1 or 2 weeks. Immunohistological studies showed the viability and integration of pericytes in the outer layer of the conduit. Mechanical tests documented a reduction in stiffness and an increase in strain at maximum load in seeded conduits in comparison with unseeded conduits. Control and pericyte-engineered conduits were then used to replace the left pulmonary artery of piglets. After 4 months, anatomical and functional integration of the grafts was confirmed using Doppler echography, cardiac magnetic resonance imaging, and histology.Conclusions These findings demonstrate the feasibility of using neonatal cardiac pericytes for reconstruction of small-size branch pulmonary arteries in a large animal model.