Kadner Alexander, Zund Gregor, Maurus Christine, Breymann Christian, Yakarisik Sidika, Kadner Gregor, Turina Marko, Hoerstrup Simon P
Clinic for Cardiovascular Surgery, University Hospital, Raemistrasse 100, CH 8091 Zurich, Switzerland.
Eur J Cardiothorac Surg. 2004 Apr;25(4):635-41. doi: 10.1016/j.ejcts.2003.12.038.
Tissue engineering of viable, autologous cardiovascular replacements with the potential to grow, repair and remodel represents an attractive approach to overcome the shortcomings of available replacements for the repair of congenital cardiac defects. Currently, vascular myofibroblast cells represent an established cell source for cardiovascular tissue engineering. Cell isolation requires the invasive harvesting of venous or arterial vessel segments prior to scaffold seeding, a technique which may not be preferable, especially in pediatric patients. This study evaluates cells isolated from human umbilical cord artery, umbilical cord vein and whole cord as alternative autologous cell sources for cardiovascular tissue engineering.
Cells were isolated from human umbilical cord artery (UCA), umbilical cord vein (UCV), whole umbilical cord (UCC) and saphenous vein segments (VC), and were expanded in culture. All three expanded cell groups were seeded on bioabsorbable copolymer strips and grown in vitro for 28 days. Isolated cells were characterized by flow cytometry, histology, immunohistochemistry, proliferation assays and compared to VC. Morphological analysis of the seeded polymer strips included histology, immunohistochemistry, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and uniaxial stress testing.
UCA, UCV and UCC demonstrated excellent cell growth properties comparable to VC. Following isolation, all three cell groups showed myofibroblast-like morphology and characteristics by staining positive for alpha-smooth muscle actin (ASMA) and vimentin. Histology and immunohistochemistry of seeded polymers showed good tissue and extracellular matrix formation containing collagen I, III and elastin. TEM showed viable myofibroblasts and the deposition of collagen fibrils and progressive growing tissue formation, with a confluent surface, was observed in SEM. No difference was found among the mechanical properties of UCA, UCV, UCC and VC tissue engineered constructs.
Tissue engineering of cardiovascular constructs by using UCA, UCV and UCC is feasible in an in vitro environment. Cell growth, morphology, characteristics and tissue formation were comparable between UCA, UCV, UCC and VC. UCC represent an attractive, readily available autologous cell source for cardiovascular tissue engineering offering the additional benefits of utilizing juvenile cells and avoiding the invasive harvesting of intact vascular structures.
构建具有生长、修复和重塑潜力的有活力的自体心血管替代物用于组织工程,是克服现有先天性心脏缺陷修复替代物缺点的一种有吸引力的方法。目前,血管肌成纤维细胞是心血管组织工程中已确立的细胞来源。细胞分离需要在支架接种前侵入性获取静脉或动脉血管段,这种技术可能并非首选,尤其是在儿科患者中。本研究评估从人脐动脉、脐静脉和整条脐带中分离的细胞作为心血管组织工程的替代自体细胞来源。
从人脐动脉(UCA)、脐静脉(UCV)、整条脐带(UCC)和大隐静脉段(VC)中分离细胞,并在培养中进行扩增。将所有三个扩增的细胞组接种到可生物吸收的共聚物条上,并在体外培养28天。通过流式细胞术、组织学、免疫组织化学、增殖测定对分离的细胞进行表征,并与VC进行比较。对接种的聚合物条进行形态学分析,包括组织学、免疫组织化学、十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳、透射电子显微镜(TEM)、扫描电子显微镜(SEM)和单轴应力测试。
UCA、UCV和UCC表现出与VC相当的优异细胞生长特性。分离后,所有三个细胞组通过α - 平滑肌肌动蛋白(ASMA)和波形蛋白染色呈阳性,显示出肌成纤维细胞样形态和特征。接种聚合物的组织学和免疫组织化学显示含有I型、III型胶原蛋白和弹性蛋白的良好组织和细胞外基质形成。TEM显示有活力的肌成纤维细胞,在SEM中观察到胶原纤维的沉积和逐渐生长的组织形成,表面融合。在UCA、UCV、UCC和VC组织工程构建体的力学性能之间未发现差异。
使用UCA、UCV和UCC进行心血管构建体的组织工程在体外环境中是可行的。UCA、UCV、UCC和VC之间的细胞生长、形态、特征和组织形成相当。UCC是心血管组织工程中一种有吸引力的、易于获得的自体细胞来源,具有利用幼年细胞和避免侵入性获取完整血管结构的额外优势。