Is anyone familiar with culturing of neonatal cardiomyocytes?
I have successfuly isolated and cultured primary neonatal cardiomyocytes from new born mice (2 days old), I would like to know how many passages can we keep them? as they are sensitive. And what is the viability after deep freezing in liquid nitrogen?
Thanks
good effort
please keep in consideration this is normal cells with the all characteristics of normal tissuebehavior. that means the number of passage not exceed 4 to 6 times. also preserve the cells with small number of passage as you can in vapour phase of liquid nitrogen. it should be with good viability for about one year if the preservation condition of liquid nitrogen tank is good.
also the number of preserved cells in cryovials should be more than 5 X 106 cell/ ml and resuscitated in small baby tissue culture flask
BR.,
Thank you, Dr. Saad, for your informative answer
I am just wondering if this instruction is common for any primary cells or also can be applied for primary cardiomyocytes cell which looks more sensitive than other cells.
Have you applied this on primary neonatal cardiomyocytes cells
I mean keeping cells more than 4 passages and preserve them in liquid nitrogen for a long time.
I frequently isolate beating cardiomyocytes from newborn rats, and unfortunately we are not able to make passages in culture. Indeed, the beating capacity of the cardiomyocytes is lost after the first passage. Have you noticed that in your culture?
Regards,
H PEYRET
I am working with different types of primary cultures from different organs liver, skin, kidney and spleen. Each types of culture has it's own behavior and characteristics even the same cell origin from different sources.
but I don't working with primary cardiomyocytes and I know this types of cells is very sensitive.
Regards,
saad
Hi Hélène Peyret
Thank you for answering.
Yes, I noticed that in my cells, beating capacity is getting less at the time when they reach confluency. It might be better to harvest neonatal cardiomyocytes immediately after getting confluent.
Best regards
Isehaq
Yes I agree with you. Moreover, neonatal cardiomyocytes don't have a high proliferation capacity. After 2 or 3 passages, I think there are almost only fibroblasts in the culture, which are less sensitive and proliferate more rapidly. This should be verified by immunofluorescence (troponin I or T). In my lab, we use cardiomyocytes only 4 to 10 days after the first seeding and don't make any passage. Good luck !
Thank you for sharing your experience and advice.
Best regards
Isehaq
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