I have tried this digestion method a few times, but a mass amount...

颖婷 王
Nov 8, 2021
protocol Protocol: Isolation of Murine Primary Aortic Smooth Muscle Cells
I have tried this digestion method a few times, but a mass amount of vsmc dies when the digestion time exceeds over 2 hours. When incubated overnight, there are just cell fragments instead of complete healthy cells under the telescope. I was wondering how the author can seperate the advent Read more Read less more less
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俊平 李 Answered Nov 10, 2021

中山大学附属第二医院

I am very glad to see your message. We are alumni. We can communicate about it. sxlijunping@163.com
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Juan Rodríguez Vita Author Answered Nov 10, 2021

CIPF

The separation of the adventitia, when done properly, does not generate too much damage. You have to consider that the media is a very elastic tissue that absorbs most of the screeching. How do you detect dead cells? How do you know that the dead cells that you observe in the incubation are VSMCs? Do you perform any kind of staining to determine the nature of these fragments? This digestion step is mainly going to dissolve the collagen fibers within the media so that VSMCs can migrate out of the aorta ring. Therefore, a lot of debris is observed but this is mainly collagen fibers.
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