I have problem last 1 month about adhesion. I am using %10 FBS, %1 pen-strep, %0.7 amphotericin and ımdm for CFPAC1. %10 FBS %2.5 HORSE SERUM ,again as much pen-strep and amphotericin DMEM for Mia-Paca 2. %10 FBS %1 PEN-STREP AND%0.7 Amphotericin and MEM for HEk 293 . I am waiting more than 2 weeks and cells are still floating in flask. Where is the problem is there any solution about that . BTW FBS and horse serum has been inactived by myself 1 hour 57 C°.
The culture method looks well. I think a problem can be the viability of the cultured cells. Some months ago, I had the same problem with the RAW 264.7 line, and the problem was its permanence in cryopreservation at -80ºC: the viable cells unfreezed concentration were very low, and if they are not viable, they will not be attached after 24 hours. So, I recommend to test the viability of cell line before the culture. Typically, they will not take a longer time to attach to the flask.
Thank you for comment Christian Segura.
I solve problem. . You were right there is problem about cell lines. We changed cell lines and everything work great.
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