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2.4K Views
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11:35 min
December 9th, 2022
DOI :
10.3791/64557-v
Chapters
0:04
Introduction
0:35
Isolation of the Gracilis Muscles
1:26
Isolation of the Tibialis Anterior (TA) and the Extensor Digitorum Longus (EDL) Muscles
3:17
Isolation of the Gastrocnemius (GA) and Soleus Muscles
4:27
Isolation of the Triceps Muscles
5:07
Isolation of the Masseter Muscles
6:08
Isolation of the Diaphragm Muscle
7:20
Muscle Digestion to a Single‐Cell Suspension
8:40
Results: Purification, Purity Assessment, and Quantitative Measurement of Quiescent Stem Cell Populations from Individual Skeletal Muscles
10:47
Conclusion
Transcript
该协议意义重大,因为它能够研究不同骨骼肌的干细胞功能,加深我们对它们如何促进体内平衡、再生和疾病进展的理解。使用该协议,我们可以从特定肌肉中分离出两个活干细胞群,并研究它们是如何工作的。要开始肌肉隔离,请用乙醇喷洒安乐死的小鼠,并将其腹部朝上放置。
用剪刀在腹部皮肤上做一个0.5厘米长的水平切口。用双手拉动皮肤,使躯干以及下肢直到后肢。找到腹股或大腿内侧肌肉,然后用一对弯曲的镊子抓住它并稍微抬起肌肉。
用剪刀做一个0.5厘米的切口,切出肌细肌。在另一条腿上重复该过程
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Summary
该协议描述了从小鼠的单个骨骼肌中分离肌肉干细胞和纤维脂肪祖细胞。该方案涉及单肌肉解剖、通过荧光活化细胞分选分离干细胞、通过免疫荧光染色进行纯度评估以及通过 5-乙炔基-2'-脱氧尿苷掺入测定法定量测量 S 期进入。
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