Kindly let me know the recent protocol for the differentiation of human adipose derived stem cells to cardiomyocytes
Hello
I suggest using differentiation of Adipose Tissue-Derived Mesenchymal Stem Cells Into Cardiomyocytes , in this protocol the Cells from the fifth passage were detached with trypsin,counted and replated at a concentration of 1x104 cells, maintained with culture media DMEN enriched with FBS 10% in 6-well plates covered by 22 mm diameter coverslips (Sarstedt) in the incubator . Twenty-four hours later, the culture media was removed,the cells washed with PBS solution, and the plates submitted to distinct protocols of cardiogenic differentiation . This process was carried out three times . Plates were incubated in a 5% CO2 incubator at 37ºC for 15 days.
pleas see attachment , i hope help you
Good Luck
Sorry to crack the news but it has been shown that mesenchymal stem cells, including adipose-derived are not very good in differentiating in cardiomyocytes. I know this is not what you want to hear but might save you tons of time.
Hi,
I agree with Jivko, efficiency is low and in in vivo even lower.
Hi, I agree, mesenchymal stromal cells, and ADSC, are notoriously bad in getting into cardiomyocytes. This has do do with the early separation of somites in the embryonic development. Expression of one or the other marker, does not mean they are truly functional cardiomyocytes, Your are better of with human embryonic stem cells, or iPS, best wishes Michael Raghunath
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