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Isolation of cardiac myocytes.. - (Sep/29/2008 )

Has anyone isolated cardiac myocytes without using a langendorff perfusion system. We are working with human cardiac myocytes and mice. We would like to do both w/o perfusion so that the isolation methods are identical between models (ie: we get artial/vent strips from human heart that can not be perfused).

Has anyone had luck in isolating myocytes using a chunk method?

We have tried a number of different things w/o much success. I either get a group of over digested myocytes that I can not make Ca++ tollerant, or I get a poor digestion where many myocytes are clumped together. Any ideas, suggestions and protocols are welcome.




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