Kitahara Hiroto, Yagi Hiroshi, Tajima Kazuki, Okamoto Kazuma, Yoshitake Akihiro, Aeba Ryo, Kudo Mikihiko, Kashima Ichiro, Kawaguchi Shinji, Hirano Akinori, Kasai Mio, Akamatsu Yuta, Oka Hidetoshi, Kitagawa Yuko, Shimizu Hideyuki
Department of Cardiovascular Surgery, Keio University School of Medicine, Tokyo, Japan.
Department of Surgery, Keio University School of Medicine, Tokyo, Japan
Interact Cardiovasc Thorac Surg. 2016 May;22(5):571-9. doi: 10.1093/icvts/ivw022. Epub 2016 Feb 21.
One of the final treatments for end-stage heart failure is heart transplantation. However, a shortage of donor hearts has created a long waiting list and limited benefits. Our ultimate goal is to create a whole beating heart fabricated on an organ scaffold for human heart transplantation. Here, we successfully performed the first transplantation using a decellularized whole porcine heart with mesenchymal stem cells.
A porcine heart was harvested following cardiac arrest induced by a high-potassium solution and stored at -80°C for 24 h. The porcine heart was completely decellularized with 1% sodium dodecyl sulphate and 1% Triton X-100 under the control of perfusion pressure (100 mmHg) and maintained at 37°C. A decellularized whole-heart scaffold was sterilized with gamma irradiation. Cultured mesenchymal stem cells were collected and either infused into the ascending aorta or injected directly into the left ventricular wall. Finally, recellularized whole-heart scaffolds were transplanted into pigs under systemic anticoagulation treatment with heparin. Coronary artery angiography of the transplanted heart graft was performed.
In our decellularization method, all cellular components were removed, preserving the heart extracellular matrix. Heterotopic transplantations were successfully performed using a decellularized heart and a recellularized heart. The scaffolds were well perfused, without bleeding from the surface or anastomosis site. Coronary angiography revealed a patent coronary artery in both scaffolds. The transplanted decellularized heart was harvested on Day 3. Haematoxylin and eosin staining showed thrombosis in the coronary arteries and migrated inflammatory cells. Haematoxylin and eosin staining of the transplanted recellularized heart showed similar findings, with the exception of injected mesenchymal stem cells.
To the best of our knowledge, this is the first report of heterotopic transplantation of a decellularized whole porcine heart with mesenchymal stem cells. The scaffolds endured surgical procedures. We detected short-term coronary artery perfusion in the transplanted scaffolds by angiography. Future studies should analyse the histological features of transplanted decellularized scaffolds and optimize the system for recellularization to apply this unique technology clinically.
心脏移植是终末期心力衰竭的最终治疗手段之一。然而，供体心脏短缺导致等待名单冗长且受益有限。我们的最终目标是制造一个在器官支架上构建的完整跳动心脏用于人体心脏移植。在此，我们成功地使用脱细胞全猪心脏与间充质干细胞进行了首次移植。
通过高钾溶液诱导心脏骤停后获取猪心脏，并在-80°C下储存24小时。在灌注压力（100 mmHg）控制下，用1%十二烷基硫酸钠和1% Triton X-100将猪心脏完全脱细胞，并维持在37°C。脱细胞全心脏支架用伽马射线灭菌。收集培养的间充质干细胞，要么注入升主动脉，要么直接注射到左心室壁。最后，将重新细胞化的全心脏支架在肝素全身抗凝治疗下移植到猪体内。对移植心脏移植物进行冠状动脉造影。
在我们的脱细胞方法中，所有细胞成分均被去除，保留了心脏细胞外基质。使用脱细胞心脏和重新细胞化心脏成功进行了异位移植。支架灌注良好，表面或吻合部位无出血。冠状动脉造影显示两个支架中的冠状动脉均通畅。移植的脱细胞心脏在第3天被取出。苏木精-伊红染色显示冠状动脉内有血栓形成且有炎性细胞迁移。移植的重新细胞化心脏的苏木精-伊红染色显示了类似结果，但注入的间充质干细胞除外。
据我们所知，这是关于脱细胞全猪心脏与间充质干细胞异位移植的首次报告。支架经受住了手术操作。我们通过血管造影检测到移植支架中的短期冠状动脉灌注。未来的研究应分析移植脱细胞支架的组织学特征，并优化重新细胞化系统以便将这项独特技术应用于临床。
