I am trying to adapt Sf21 cells to serum-free adherent culture in Gibco® Sf-900™ III SFM. According to the manufacturer protocol, cells will be attached tightly and it will require additional force. Unfortunately, in my case the cells are attached too tightly and every passage I lose cells. Also many cells become in elongated shape after adding 75% Sf-900™ III SFM. I have to solve this problem.
I'm asking for kind advice from experts. Thank you so much for considering your very precious time.
keep them on a frozen ice pack for 4-5 mins and then flush them with micropipette using the same media inside the flask. The cold cells cant bind the wall. So the binding will be loosened and will come to the medium easily. But do not keep them more than 5mins. It might give a cold shock to them
Shayanki Lahiri Charles D Anderson
Thank you so much for your kind advice. Besides placing into 4 degree, I'm letting them to grow up to 100% of the confluency level. Now it seems like it's OK. Anyway, I'm still worried about the elongated cells.
I am also curious about the elongated cells, I am growing sf9 cells and I see these elongated cells and am wondering if they are just the cells or are contamination. Any advice is appreciated!!
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