Medicine
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几内亚猪模型心肌损伤后 hiPSC-derived 衍生心肌贴片的植入
Chapters
Summary March 18th, 2019
Please note that all translations are automatically generated.
Click here for the English version.
在这里, 我们提出了一个方案, 诱导左心室内皮损伤, 然后植入心肌贴片, 从人类 ips 细胞心肌细胞在豚鼠模型中获得。
Transcript
该方法的目的是评估工程心脏组织植入小动物冷冻治疗模型的功能后果。这种方法的优点是，它允许诱导相对可重复的心肌梗塞与稳定的梗塞大小。在确认对头趾捏没有反应后，将麻醉的豚鼠放在40至42摄氏度的加热平台上。
涂抹眼药膏，剃光，脱皮胸部。将 25 摄氏度超声波传感器凝胶涂抹在外露的皮肤上，将超声心动图系统的传感器放置在胸部，以 B 模式从右颈部向主主阀平面的左腿获取二维副星长轴视图，同时显示左心室顶点，以调查术前左心室功能。然后将传感器转动 90 度，在中轴级别获得短轴 B 模式视图。
获得最后一幅图像后，将豚鼠转移到手术台上，将四肢固定在展开的鹰位置。用两个连续的碘基和80%乙醇磨砂对裸露的皮肤进行消毒，并在气管区域进行1.5厘米的垂直切口。模糊解剖覆盖气管的肌肉。
当可以观察到气管时，用 18 量表 IV 管刺穿空气管，并将气管的柔性部分插入气管。将气管连接到动物呼吸器，在手术过程中进行连续通风，并计算从第一个成本间空间开始的肋骨空间，以定位第五个成本间空间。在豚鼠左侧的第五个间空间上做一个两厘米水平切口，在用电烧解剖肌肉之前切除皮下组织，直到达到成本间肌肉。
用钳子和小剪刀轻轻解剖成本间肌肉，直到达到胸膜空间，左肺可以可视化。在肋骨之间插入一个缩回器，并小心地打开缩回器，直到获得心脏的美景。在左心室前壁区域打开大约一厘米的腹孔，并在诱导左心室的低温损伤时，在左肺上放置压缩，以保护肺部免受损伤。
接下来，将横截面直径为 0.5 厘米的液氮侵蚀金属邮票的尖端冷却三分钟，然后将探头压在心脏的左前壁上 30 秒。然后在邮票内放置一个200摄氏度的电焊铁，以加热邮票，使金属可以从组织中去除。第三次盖章应用后，将缩回器从腹间空间中拆下，并夹住呼吸机的流出管两秒钟，以最大压力充气肺部，避免肺部进退。
然后用两个 4 - 0 缝合关闭肋骨， 用 5 - 0 运行缝合的肋骨上的肌肉和 5 - 0 缝合皮肤。取出气管后，使用单个 8-0缝合关闭气管的穿刺点，用三个单针5-0缝合关闭伤口。冷冻伤害后七天，去除缝合线，在左侧疤痕区域做一个两厘米水平皮肤切口，并使用电烧机轻轻解剖多余的胸膜粘附。
小心地用剪刀打开胸膜空间，并插入肋骨展开器以暴露心脏。与健康的心肌相比，用淡淡的颜色直观地识别梗死区域，并在梗塞区域放置一个工程心脏组织贴片。用两个 8 - 0 保护补丁心脏非梗塞区域两侧的缝合，使肺部有压力，避免肺部进一步扩张。
然后从成本空间中卸下缩回器，然后关闭动物，如演示的。工程心脏组织植入四周后，进行跨胸超声心动图，用于评估左心室功能加班的变化。在冷冻损伤四周后，梅森三色染色显示，平均超过25%的左心室心肌有大的跨体疤痕。
肌萎缩素染色表明大型心肌移植物已部分重新肌肉化为人类Ku80的疤痕和染色，证实了新形成的心肌的人类起源。植入工程心脏组织的跨胸超时心心动图监测显示，左心室弹出分数、分数面积缩短以及左心室和舒张直径减少。在这种重新操作中，必须小心地打开胸腔腔，因为胸壁和左心室之间的粘附存在，并且有一定的风险伤害左心室心肌。
植入人体工程心脏组织后，可以使用多种方法监测左心室喷射分数的变化。除了超声心动图外，压力体积回路的侵入性测量可用于监测这些变化。这种方法允许研究人体工程组织植入的所有方面。
功能后果以及潜在的风险，如心律失常可以在这个小动物模型中进行评估。
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