I am culturing 'Human Umbilical Vein Endothelial Cells' & 'Normal Human Lung Fibroblasts' from the PELOBiotech in their 'Endothelial Cell Growth Medium' & 'Fibroblast Growth Medium'. I am wondering whether it would make a difference if I used the 'Accutase solution' instead of 'Trypsin' for cell detachment! Any experiences? (For the HUVECs, I am using Collagen-I coated plates)
Hello Fatemeh!
Both trypsin and acutase can be used to detach cells. here is the protocol and article
Article Passaging and colony expansion of human pluripotent stem cel...
hi
Don't worry, it doesn't matter.
1-For FACS assay "accutase " better than " Trypsin-EDTA "
2- For general application " Trypsin-EDTA " stronger & cheaper than "accutase "
good luck
Thanks! So, no major differences! (I was just wondering since we are planning 3D culture...)
Both the two chemicals can be used to detach the cells. I usually used Trysin-EDTA to quickly detach and preserve the viability of the cells.
hello, I met the same problem. In a 3D culture system, using 0.25% trypsin for 3 minutes cannot detach well. Could you share the final effective digestion method？Thanks a lot!
Luping Shen Sure!
I ended up using the 0.5g-trypsin/0.2g-EDTA SIGMA product with a slightly modified trypsinizing protocol. I treat the cells with trypsin for about 10 seconds and, after aspirating the trypsin all out, let the plate be at room temperature for 1-2 min. Tapping on the side of the plate one-two times, I can easily collect the ECs off the collagen-coated surface.
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