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Developmental Biology
在免疫缺陷小鼠中表演人类骨骼肌异种移植物
在免疫缺陷小鼠中表演人类骨骼肌异种移植物
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
Performing Human Skeletal Muscle Xenografts in Immunodeficient Mice

在免疫缺陷小鼠中表演人类骨骼肌异种移植物

Full Text
10,513 Views
07:48 min
September 16, 2019

DOI: 10.3791/59966-v

Kyla A. Britson1,2, Aaron D. Black2,3, Kathryn R. Wagner1,2,3,4, Thomas E. Lloyd1,2,4

1Graduate Program in Cellular and Molecular Medicine,Johns Hopkins University School of Medicine, 2Department of Neurology,Johns Hopkins University School of Medicine, 3Kennedy Krieger Institute, 4Department of Neuroscience,Johns Hopkins University School of Medicine

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在传统的实验室模型系统中,复杂的人类疾病对建模可能具有挑战性。在这里,我们描述了一种通过将人类骨骼肌活检移植到免疫缺陷小鼠中来模拟人类肌肉疾病的外科方法。

该协议是重要的,因为它可以用来创建人类骨骼肌异种移植,可用于建模肌肉疾病和进行临床前治疗测试。这种异种移植模型使研究人员能够研究人体体内的肌肉,以更好地了解人类肌肉细胞生物学,并开发稀有或后天肌肉疾病的新模型。获得人体肌肉活检后,将标本放在含有肌肉介质的100至15毫米培养皿中,并使用立体显微镜和手术剪刀去除剩余的筋膜或脂肪组织。

将肌肉活检分解成大约七由三毫米的碎片,注意纤维在标本内纵向排列。接下来,将含有解剖肌肉的培养皿转移到冰上,将合成的、不可吸收的缝合线放在100乘15毫米的培养皿中,其中含有70%乙醇。在确认8至12周大的NOT-Rag-Gamma小鼠对脚趾捏没有反应后,对免疫缺陷动物的眼睛涂抹软膏,并使用修剪器将头部从脚踝到膝盖的头饰覆盖。

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