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全心脏去细胞化——优化基于经典十二烷基硫酸钠的去细胞化方案

Decellularization of Full Heart-Optimizing the Classical Sodium-Dodecyl-Sulfate-Based Decellularization Protocol.

作者信息

Al-Hejailan Reem, Weigel Tobias, Schürlein Sebastian, Berger Constantin, Al-Mohanna Futwan, Hansmann Jan

机构信息

Department of Cell Biology, King Faisal Specialist Hospital and Research Centre, Riyadh 11211, Saudi Arabia.

Department of Tissue Engineering and Regenerative Medicine, University Hospital Würzburg, 97070 Würzburg, Germany.

出版信息

Bioengineering (Basel). 2022 Apr 1;9(4):147. doi: 10.3390/bioengineering9040147.

Abstract

Compared to cell therapy, where cells are injected into a defect region, the treatment of heart infarction with cells seeded in a vascularized scaffold bears advantages, such as an immediate nutrient supply or a controllable and persistent localization of cells. For this purpose, decellularized native tissues are a preferable choice as they provide an in vivo-like microenvironment. However, the quality of such scaffolds strongly depends on the decellularization process. Therefore, two protocols based on sodium dodecyl sulfate or sodium deoxycholate were tailored and optimized for the decellularization of a porcine heart. The obtained scaffolds were tested for their applicability to generate vascularized cardiac patches. Decellularization with sodium dodecyl sulfate was found to be more suitable and resulted in scaffolds with a low amount of DNA, a highly preserved extracellular matrix composition, and structure shown by GAG quantification and immunohistochemistry. After seeding human endothelial cells into the vasculature, a coagulation assay demonstrated the functionality of the endothelial cells to minimize the clotting of blood. Human-induced pluripotent-stem-cell-derived cardiomyocytes in co-culture with fibroblasts and mesenchymal stem cells transferred the scaffold into a vascularized cardiac patch spontaneously contracting with a frequency of 25.61 ± 5.99 beats/min for over 16 weeks. The customized decellularization protocol based on sodium dodecyl sulfate renders a step towards a preclinical evaluation of the scaffolds.

摘要

与将细胞注射到缺损区域的细胞疗法相比,用接种在血管化支架中的细胞治疗心肌梗死具有诸多优势,比如能立即提供营养供应,或者使细胞实现可控且持久的定位。为此,脱细胞的天然组织是更为理想的选择,因为它们能提供类似体内的微环境。然而,此类支架的质量在很大程度上取决于脱细胞过程。因此,针对猪心脏的脱细胞处理,定制并优化了两种基于十二烷基硫酸钠或脱氧胆酸钠的方案。对所得支架进行了生成血管化心脏补片适用性的测试。结果发现,用十二烷基硫酸钠进行脱细胞处理更为合适,所得到的支架DNA含量低,细胞外基质成分高度保留,通过糖胺聚糖定量和免疫组织化学分析显示出其结构。将人内皮细胞接种到脉管系统后,凝血试验证明了内皮细胞具有使血液凝固最小化的功能。与成纤维细胞和间充质干细胞共培养的人诱导多能干细胞衍生的心肌细胞,使支架自发转变为血管化心脏补片,在超过16周的时间里以每分钟25.61±5.99次的频率自主收缩。基于十二烷基硫酸钠的定制脱细胞方案朝着支架的临床前评估迈出了一步。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7282/9032179/ea10b2a2d9c3/bioengineering-09-00147-g001.jpg

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