Using the recommended media and temperature to grow the cells but they die following the first sub-culture. When revived from frozen they look very healthy but as soon as they are split most of the cells die. Have tried different batches of FBS to no avail. Does anyone have any ideas?
Hello ,
I think u should increase concentration of FBS in ur composed culture medium. And do manual passaging for first few subculture. When work with cryopreserved cells follow appropriate thawing protocol.
verify pH and osmolality of composed medium , before seeding the cells.
All the best.
Hi Lindsay
Can you please share the pics of your culture highlighting what problem you are facing? Because I do not find any problem in culturing and sub-culturing hFOB 1.19. The only issue I found with hFOB 1.19 is with increase in passage number, its doubling time increase else these cells grow well.
Monika
Hi Lindsay,
I do agree with previous comments, increasing passages slows down growth. I also find very important not to split the cells too hard: the recommended 1:4 ratio is too high for the first subculture after reviving from frozen.
Hope this helps,
Lucia
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