Enhanced Methods for Needle Biopsy and Cryopreservation of Skeletal Muscle in Older Adults.
Enhanced Methods for Needle Biopsy and Cryopreservation of Skeletal Muscle in Older Adults.
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DOI：
10.37421/jch.2020.11.553
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发表时间：
2020-01-01
期刊：
影响因子：
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通讯作者：
Wanagat, Jonathan
中科院分区：
文献类型：
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作者：
Lee, Cathy C;Hoang, Austin;Wanagat, Jonathan
人体肌肉活检对于诊断、研究和监测治疗试验越来越重要。我们研究了使用独立的真空辅助活检系统和新型肌肉冷冻技术来改进、简化和标准化老年人的人体肌肉活检收集和冷冻保存。 VACORA 真空辅助活检系统对 12 名年龄从 57 岁到 80 岁的个体进行了肌肉活检。这种基于办公室的方法具有良好的耐受性，因为它是微创的，仅使用局部麻醉剂，并且恢复很快。为了最大限度地提高活检样本的质量和重现性，我们开发了一种新型的肌肉样本冷冻方案。将新鲜的肌肉活检样本放入现成的组织盒中，然后直接冷冻在液氮中。在这种修改后的速冻方案之后，将冷冻的肌肉样品包埋在包埋介质中以进行冷冻切片。我们检查了这种冷冻方法在啮齿动物和人类肌肉样本的组织学切片中的效果。 VACORA 活检系统从单个活检部位提供多达四个骨骼肌核心样本。 12 名老年人的活检样本平均每份重量为 147.5 ± 11 毫克，大小和形状一致。无并发症，残留疤痕小于10毫米。使用标准组织盒在液氮中直接冷冻的冷冻方法产生了适合组织学分析的高质量冷冻肌肉组织，无需异戊烷并且几乎没有冻融损伤。这些增强功能简化并提高了我们肌肉活检方案的一致性，并为人体肌肉衰老和疾病的多维下游研究提供了足够的高质量样本。
Human muscle biopsies are increasingly important for diagnosis, research, and to monitor therapeutic trials. We examined the use of a self-contained, vacuum-assisted biopsy system and a novel muscle freezing technique to improve, simplify, and standardize human muscle biopsy collection and cryopreservation in older adults. The VACORA vacuum-assisted biopsy system was deployed in muscle biopsies of 12 individuals ranging in age from 57 to 80 years. This office-based approach was well tolerated as it is minimally invasive, uses only local anesthetic, and has a quick recovery. To maximize biopsy sample quality and reproducibility, we developed a novel muscle sample freezing protocol. Fresh muscle biopsy samples were placed into readily available tissue cassettes followed by direct freezing in liquid nitrogen. After this modified snap freezing protocol, frozen muscle samples were enrobed in embedding medium for cryosectioning. We examined the effect of this freezing approach in histological sections of rodent and human muscle samples. The VACORA Biopsy System provided as many as four skeletal muscle core samples from a single biopsy site. Biopsy samples from 12 older adults weighed an average of 147.5 ± 11 mg each and had a consistent size and shape. There were no complications, and the residual scar is less than 10 mm. The freezing method using standard tissue cassettes with direct freezing in liquid nitrogen yielded high quality cryopreserved muscle tissue suitable for histological analysis without the need for isopentane and with little to no freeze-thaw damage. These enhancements have streamlined and improved the consistency of our muscle biopsy protocol and provide sufficient high-quality sample for multi-dimensional downstream studies of human muscle in aging and disease.