Preparation of Cardiac Extracellular Matrix from an Intact Porcine Heart
Preparation of Cardiac Extracellular Matrix from an Intact Porcine Heart
复制标题
DOI：
10.1089/ten.tec.2009.0392
复制
发表时间：
2010-06-01
影响因子：
3
通讯作者：
Badylak, Stephen F.
中科院分区：
文献类型：
--
作者：
Wainwright, John M.;Czajka, Caitlin A.;Badylak, Stephen F.
整个器官工程将受益于由完整的器官特异性细胞外基质(ECM)制成的三维支架。这样的支架所提供的微环境和结构可能会支持适合现场的细胞分化和空间组织。从完整的器官中生产这种支架的方法需要定制的脱细胞方案。在本研究中，利用脉动逆行主动脉灌流，在不到10h的时间内成功地将完整的成年猪心脏脱细胞。使用酶、非离子洗涤剂、离子洗涤剂和酸性溶液与低渗和高渗冲洗液连续灌流，系统地去除细胞内容物。生成的心脏ECM保留了胶原、弹性蛋白和糖胺聚糖，并保持了机械完整性。心脏ECM支持鸡心肌细胞在体外形成有组织的肌节结构。完整的脱细胞猪心为未来的临床前研究和最终的临床应用提供了一种组织工程模板。
Whole organ engineering would benefit from a three-dimensional scaffold produced from intact organ-specific extracellular matrix (ECM). The microenvironment and architecture provided by such a scaffold would likely support site-appropriate cell differentiation and spatial organization. The methods to produce such scaffolds from intact organs require customized decellularization protocols. In the present study, intact adult porcine hearts were successfully decellularized in less than 10 h using pulsatile retrograde aortic perfusion. Serial perfusion of an enzymatic, nonionic detergent, ionic detergent, and acid solution with hypotonic and hypertonic rinses was used to systematically remove cellular content. The resultant cardiac ECM retained collagen, elastin, and glycosaminoglycans, and mechanical integrity. Cardiac ECM supported the formation of organized chicken cardiomyocyte sarcomere structure in vitro. The intact decellularized porcine heart provides a tissue engineering template that may be beneficial for future preclinical studies and eventual clinical applications.