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2.3K Views
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07:31 min
April 16th, 2021
DOI :
10.3791/62439-v
Chapters
0:04
Introduction
0:34
Tissue Preparation and Processing
1:32
Valve Endothelial Cell (VEC) Isolation, Expansion, and Confirmation
4:01
Valve Interstitial Cell (VIC) Isolation, Expansion, and Confirmation
5:40
Results: Analysis of Isolated Human Primary Valve Cells
6:47
Conclusion
Transcript
分离人原代瓣膜内皮和间质细胞对于了解钙化性主动脉瓣疾病发病机制的潜在机制至关重要。当瓣膜从天然组织中切除时,细胞活力下降。因此,我们确定了一种延长细胞活力的方法。
收到提取的瓣膜组织样本后,提取主动脉根部并将组织浸入含有无菌冲洗溶液的50毫升锥形管中。在摇臂上的冰桶中孵育10分钟后,用70%乙醇喷洒试管。在无菌组织培养罩中,将组织转移到培养皿中并切除两个瓣膜小叶。
将一张小册子放入低温小瓶中,并在液氮中快速冷冻组织以零下80摄氏度储存。为了固定组织以进行钙含量染色,
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Summary
该协议描述了在外科主动脉瓣置换手术期间或从尸体组织中提取的人主动脉瓣的收集,以及随后对患者特异性原发性瓣膜内皮和间质细胞的分离、扩增和表征。其中包括有关确保细胞活力和表型特异性所需过程的重要细节。
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