I'm working with SiHa cell line. Every time I subculture it, I have difficulty getting the cells detached. I use a 0.025% trypsin-.037% EDTA solution.
Try washing the cells, before use trypsin, with a PBS without calcium and cloridium to remove calcium and semplify cells detachment.
@ Valeria : I did try that too. Doesnt seem to make much of a difference :(
Yes. In the incubator. I take the flask out and view it under the microscope. The cells are always rounded up but after inactivating the trypsin and even after tapping the flask they do not come off.
First of all, wash the cell well by PBS after you throw away the media, then add Trypsin and digest in the 37 degree incubator for 3 to 5 minutes. After that, tap the dish gently and leave the trypsin inside and add fresh media together. then pipette it up and down. So even tough cells you can detach it. Good luck.
Hi Lakhsmi, could you send me your mail id so that I can send you the paper? I am not sure how to send in researchgate. Mine is partha1kundu(at)gmail.com
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