Hi all,
Just out of curiosity, is there any ways to culture cell line which goes from suspension to adherent type?
I am actually culturing fish spleen cells however, it does not appear attached to bottom as referred protocol, I read somewhere bcos of its hematopoetic properties(?), is there anyone that can share any answer?
Thank you.
Hello, Tang.
I have no experience in fish spleen cells, however, you should follow all the procedures exactly to compare the previously reported one. The stage of developmental animals, male or female, or are you using the same fish species etc. Otherwise, reproducible results could not be obtained. If you are exploring in other fish, then use fry or young, juvenile fish. If it wouldn't work, coculture with endothelial cells from young fish.
Best wishes.
which cell culture media you are using? for anchorage you need serum (preferably 10% Fetal Calf Serum) in your cell culture media.
Hello Lee,
Yes i am using same fish species and same protocol as reported, however, it does not work as i expected. Thus, I am thinking whether i miss out on something or there is different culturing methods
Hi Samad,
I am using Leibovitz-15 medium for cell culture with 20% fetal bovine serum for first passage and protocol mentioned i can change to 10% FBS after i got the monolayer growing but i cant see any cell growing after fifth day culturing
Hi, Tang.
We used to culture primary embryonic cells isolated from zebrafish. A right L-15 medium for an atmosphere of air and temperature (~28C) were major differences for them, I recall. Is the L-15 medium type right one? Gas condition for fish in the air (should goes with medium type together) and temperature.
Good luck.
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