Performing Human Skeletal Muscle Xenografts in Immunodeficient Mice
Performing Human Skeletal Muscle Xenografts in Immunodeficient Mice
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DOI：
10.3791/59966
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发表时间：
2019-09-01
影响因子：
1.2
通讯作者：
Lloyd, Thomas E.
中科院分区：
文献类型：
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作者：
Britson, Kyla A.;Black, Aaron D.;Lloyd, Thomas E.
在动物研究中观察到的治疗效果通常无法在临床试验中重现。虽然这个问题是多方面的，但失败的一个原因是使用了不适当的实验室模型。在传统的实验室生物体中模拟复杂的人类疾病是具有挑战性的，但这个问题可以通过研究人类异种移植来规避。我们在这里描述的手术方法允许创建人骨骼肌异种移植物，其可用于模拟肌肉疾病并进行临床前治疗测试。根据机构审查委员会（IRB）批准的方案，从患者获得骨骼肌样本，然后移植到NOD-Rag 1（null）IL 2 r γ（null）（NRG）宿主小鼠中。这些小鼠是移植研究的理想宿主，因为它们不能产生成熟的淋巴细胞，因此不能产生细胞介导的和体液适应性免疫应答。用异氟烷麻醉宿主小鼠，并去除小鼠胫骨前肌和趾长伸肌。然后将一块人体肌肉放置在空的胫骨间室中，并缝合到腓骨长肌的近端和远端肌腱上。异种移植的肌肉由小鼠宿主自发地血管化和神经支配，从而产生可用作临床前研究的模型的健壮再生的人类肌肉。
Treatment effects observed in animal studies often fail to be recapitulated in clinical trials. While this problem is multifaceted, one reason for this failure is the use of inadequate laboratory models. It is challenging to model complex human diseases in traditional laboratory organisms, but this issue can be circumvented through the study of human xenografts. The surgical method we describe here allows for the creation of human skeletal muscle xenografts, which can be used to model muscle disease and to carry out preclinical therapeutic testing. Under an Institutional Review Board (IRB)-approved protocol, skeletal muscle specimens are acquired from patients and then transplanted into NOD-Rag1(null) IL2r gamma(null) (NRG) host mice. These mice are ideal hosts for transplantation studies due to their inability to make mature lymphocytes and are thus unable to develop cell-mediated and humoral adaptive immune responses. Host mice are anaesthetized with isoflurane, and the mouse tibialis anterior and extensor digitorum longus muscles are removed. A piece of human muscle is then placed in the empty tibial compartment and sutured to the proximal and distal tendons of the peroneus longus muscle. The xenografted muscle is spontaneously vascularized and innervated by the mouse host, resulting in robustly regenerated human muscle that can serve as a model for preclinical studies.