Many children born with congenital heart disease need a heart valve repair or replacement. Currently available repair materials and valve replacements are incapable of growth, repair, and adaptation, rendering them inadequate for growing children. Heart valve tissue engineering (HVTE) aims to develop living replacement valves that can meet these needs. Among numerous cell sources for in vitro HVTE, umbilical cord perivascular cells (UCPVCs) are particularly attractive because they are autologous, readily available, and have excellent regenerative capacity. As an essential step toward preclinical testing of heart valves engineered from UCPVCs, the goal of this study was to establish methods to isolate, expand, and promote extracellular matrix (ECM) synthesis by UCPVCs from pigs (porcine umbilical cord perivascular cells [pUCPVCs]), as a relevant preclinical model. We determined that Dulbecco's modified Eagle's medium with 20% fetal bovine serum supported isolation and substantial expansion of pUCPVCs, whereas media designed for human mesenchymal stromal cell (MSC) expansion did not. We further demonstrated the capacity of pUCPVCs to synthesize the main ECM components of heart valves (collagen type I, elastin, and glycosaminoglycans), with maximal collagen and elastin per-cell production occurring in serum-free culture conditions using StemMACS™ MSC Expansion Media. Altogether, these results establish protocols that enable the use of pUCPVCs as a viable cell source for preclinical testing of engineered heart valves.

中文翻译：

---

猪脐带血管周围细胞用于组织工程心脏瓣膜的临床前测试

许多先天性心脏病患儿需要心脏瓣膜修复或更换。目前可用的修复材料和瓣膜替代物无法生长、修复和适应，使其不适合成长中的儿童。心脏瓣膜组织工程 (HVTE) 旨在开发能够满足这些需求的活体置换瓣膜。在众多体外细胞来源中HVTE、脐带血管周围细胞 (UCPVC) 特别有吸引力，因为它们是自体的、容易获得的并且具有出色的再生能力。作为对由 UCPVCs 设计的心脏瓣膜进行临床前测试的重要一步，本研究的目的是建立分离、扩展和促进猪 UCPVCs（猪脐带血管周围细胞 [pUCPVCs]）合成细胞外基质 (ECM) 的方法，作为相关的临床前模型。我们确定，含有 20% 胎牛血清的 Dulbecco 改良 Eagle 培养基支持 pUCPVC 的分离和大量扩增，而设计用于人间充质基质细胞 (MSC) 扩增的培养基则不支持。我们进一步证明了 pUCPVCs 合成心脏瓣膜主要 ECM 成分（I 型胶原蛋白、弹性蛋白、和糖胺聚糖），在使用 StemMACS™ MSC 扩增培养基的无血清培养条件下，每个细胞产生最大的胶原蛋白和弹性蛋白。总而言之，这些结果建立了协议，使 pUCPVC 能够作为可行的细胞来源用于工程心脏瓣膜的临床前测试。