Medicine
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建立干细胞治疗心肌梗塞心力衰竭的猪模型
Chapters
Summary May 25th, 2020
Please note that all translations are automatically generated.
Click here for the English version.
我们寻求建立一个猪模型的心力衰竭引起的左环弹性动脉阻塞和快速起搏,以测试心肌内给给干细胞的效果和安全性的细胞为基础的疗法。
Transcript
这种猪心脏衰竭模型,由左环状动脉阻塞和快速起搏引起的,可用于评估直接心肌内干细胞注射对心脏再生的影响。我们的猪群心力衰竭模型相当稳定。该模型的新梗塞尺寸约为左心室的 16%,左心室注射梗塞从基线至少减少 40%。
将实验猪固定在超生位置,在胡萝卜三角中定位右胡萝卜动脉和杂耍静脉,并使用止血钳和无菌技术分离右胡萝卜动脉和血管。将右胡萝卜动脉和杂耍静脉的端部拉开,并使用血管瘤来调节右杂耍静脉。在 X 射线指导下将心脏起搏器铅插入右心室后,使用钳子分离胸肌和前部骨质肌,并将心脏起搏器连接到铅。
然后在两块肌肉之间植入心脏起搏器。然后用 2 - 0 的丝绸关闭两块肌肉。要评估左心室功能的变化,请隔离股骨三角形中的右股动脉和股骨静脉,并计算右股动脉与血管瘤。
通过血管将导线放入股骨静脉,取出血管。在导线的指导下,将 12 个法国护套插入股骨静脉并拆下导丝。可以用九个法国护套来调节右股动脉,正如刚刚证明的。
并在 X 射线指导下,通过 12 个法国护套插入气球导管进入内部 Vena cava。使用压力体积信号处理器在同位素盐水中校准 7 个法国压力体积导管,并在 X 射线指导下通过 9 个法国护套将导管插入左心室顶点。暂停通风并使用压力体积信号处理器测量左心室最大和负压导数,以及收缩压力和结束舒张压力。
然后重新启动通风。对于心肌梗塞诱导,在计算右心肌动脉与八个法国护套之前,在计算右心肌动脉之前,在一小时内静脉注射5毫克每公斤阿米奥达龙和每公斤1.5毫克利多卡因玻利瓦尔,以防止心室心律失常。通过六根法国护套进行冠状动脉造影,通过标准 C 臂荧光镜引导的放置护套进行。
在X射线指导下,左环状冠状动脉向第一个迟钝边缘分支闭塞,皮下透骨血管成形术扩张气球导管膨胀。将1毫升700微米的海绵微球与7毫升的 iohexol 混合在10毫升注射器中,然后通过气球导管注射混合物,以阻止左环状冠状动脉。然后放气气球,并执行造影检查以确认遮挡。
梗死诱导负荷8周后,将两次10至第八个人类诱导多能干细胞衍生的中流体干细胞在两毫升正常盐水中放入5毫升注射器中,并在麻醉实验动物的顶点节拍区域周围消毒10厘米。使用缩回器在四到五个间空间进行左胸切除术,并进行心内切除术以暴露梗死的横向壁。当非法区域可见时,每次注射进行5至8次心肌注射,注射约300微升细胞,然后用铁丝关闭间空间,用2-0丝线关闭肌肉层,用2-0维丽线关闭皮下组织和皮肤层。
为了评估细胞治疗后心室性,通过股骨静脉将六个法国电生理导管插入右心室顶点,并在电生理记录系统中以200毫米/秒的速度显示心内记录,并显示表面心电图leas I、II和III。使用刺激器以两倍于舒张阈值的两毫秒脉冲宽度提供,并在一个 200 毫秒和一个 300 毫秒的驱动周期长度下提供八个刺激的起搏列车,然后是一两个过早的额外刺激。然后按顺序缩短耦合间隔,直到心室有效耐火性心律失常周期被诱导,注意到存在一种不可诱导的持续心室性心律失常。
在这个具有代表性的实验中,连续超声心动图检查表明,左心室弹射分数从基线显著减少,而左心室末舒张维度和末体收缩量在心肌梗塞诱导后八周内显著增加。相比之下,左心室弹射分数显著增加,细胞移植组的末收缩量在给给后八周显著下降。负压导数和末收缩压体积关系在心肌梗塞诱导后八周内从基线显著下降,而心肌内肌细胞干细胞的内向作用导致这些功能增加。
从每只动物的五到七个连续一厘米厚的部位测量左心室内壁厚度,发现左心室梗塞的百分比约为16% 移植后8周的注射部位周围没有细胞存活。但是,在近脑外区域可以看到少量幸存的注射干细胞。在有心力衰竭的动物中,可引致持续心室性心律失常的事故很容易增加。
值得注意的是,干细胞移植没有显著改变基础心肌底,以减少心室性。心室性大出血症。由于左环弯曲冠状动脉阻塞可能诱发心律失常。请务必监测心电图,并立即使用外部双相除颤器,以重建鼻窦。
该方法为测试干细胞疗法的疗效提供了稳定且可重复的临床相关大型心力衰竭动物模型。
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