Tee Richard, Morrison Wayne Allan, Dilley Rodney J
O'Brien Institute, 42, Fitzroy Street, Fitzroy, Victoria, 3065, Australia.
Department of Surgery, University of Melbourne, Victoria, 3065, Australia.
Microsurgery. 2018 Jul;38(5):544-552. doi: 10.1002/micr.30325. Epub 2018 Apr 6.
The survival of engineered cardiac muscle 'grafts' to the epicardium is limited by vascularization post-transplantation in rat models. In this article, we describe the methodology of a novel rat model that allows for the transplantation of an engineered cardiac muscle flap (ECMF) onto the epicardium.
A total of 40 rats were used. Twenty-four neonatal rats were used to harvest cardiomyocytes. At week 1, ECMF were generated by seeding cardiomyocytes into the arteriovenous loop (AVL) tissue engineering chamber implanted into the right groin of adult rats (n = 8). At week 6, the ECMF were harvested based on a pedicle along the femoral-iliac-abdominal vessel and anastomosed to the neck vessels of the recipient syngeneic adult rats (n = 8). The flaps were delivered into the thoracic cavity and onto the epicardium. The transplanted flaps were harvested at week 10. Survival of the flaps was assessed by the patency of anastomoses and viability of the cardiomyocytes through histological analysis (hematoxylin and eosin [H&E], desmin, and von Willebrand factor [vWF] immunostaining).
Six out of 8 rats survived the transplantation procedure. These remaining 6 recipient rats survived until harvest time point at 4 weeks post-transplantation. The mean area of the flap was 46.7mm . Six out of 6 flaps harvested at week 10 showed viable cardiomyocytes using desmin immunostaining and vascular channels were seen at the interface between flap and epicardium.
This is a technically feasible model that will be useful for future assessment of different cardiac stem cell implants and their functional significance in rat heart models.
在大鼠模型中，工程化心肌“移植物”在移植到心外膜后，其存活受到移植后血管化的限制。在本文中，我们描述了一种新型大鼠模型的方法，该模型允许将工程化心肌瓣（ECMF）移植到心外膜上。
共使用40只大鼠。24只新生大鼠用于获取心肌细胞。在第1周，通过将心肌细胞接种到植入成年大鼠右腹股沟的动静脉环（AVL）组织工程腔室中来生成ECMF（n = 8）。在第6周，基于沿股-髂-腹血管的蒂收获ECMF，并将其与同基因成年受体大鼠（n = 8）的颈部血管吻合。将瓣片送入胸腔并置于心外膜上。在第10周收获移植的瓣片。通过吻合口的通畅情况以及通过组织学分析（苏木精和伊红[H&E]、结蛋白和血管性血友病因子[vWF]免疫染色）评估心肌细胞的活力来评估瓣片的存活情况。
8只大鼠中有6只在移植手术后存活。其余6只受体大鼠存活至移植后4周的收获时间点。瓣片的平均面积为46.7平方毫米。在第10周收获的6个瓣片中，有6个通过结蛋白免疫染色显示心肌细胞存活，并且在瓣片与心外膜的界面处可见血管通道。
这是一个技术上可行的模型，将有助于未来评估不同心脏干细胞植入物及其在大鼠心脏模型中的功能意义。
