What precautions one should take to maintain viability of stem cells of adipose tissue, during their isolation
You need to work in GMP conditions and freezing and thawing in accordance of literature consensus! At first, you need attention to velocity of temperature decrease and second, you need attention to temperature increasing with cryoprotector, like DMSO and needs to do fast thawing. DMSO is cytotoxic in ambient temperature. The adipose derived stem cells have not difficulties for cryopreservation, but maybe the adherent markers could be reduced after these proceedings and interfere in adhesion and the viability will be maintained. See attached.
Hello Nisha,
Completely agree with Amita. In addition to the above suggestions, take care not to centrifuge stem cells for too long during pellet down step of single cell suspension you finally get after digestion and even avoid using high centrifugation speeds. It applies to upscaling your culture also. Generally stem cells settle down very nicely for 2k rpm for 2 minutes. High speed centrifugation for longer time is definitely going to kill cells. Last but not least, don't do harsh pipetting on cells, try to handle them gently like babies. They will respond well to your love and care.
All the best !
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