I have cultured HPDE cells twice. They were growing good enough and when confluent, I splitted them once with Trypsin and acutase separately but they are almost all dead and didnt attach to the surface.
Does somebody has experience or recommendations?
Amanda O'Donnell
Have you tried using trypsin inhibitor (soy-bean) after they have detached, then spinning down to remove the trypsin?
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Yong Weon Yi
Are your HPDE cells normal primary cells? If so, it would be better to avoid confluent growth of cells before subculture. Normal human cells, such as HUVEC, should be subcultured before confluent.
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Shakhawan Mustafa
No Amanda, I didnt remove the trypsin but inactivated it with serum.
Yong Weon Yi, they were not 100% confluent when I trypsinized them.
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