Hey guys, just some background information first:
Growing H9C2 cells and trying to ensure they maintain the phenotye of Cardiac cells. They are grown in T150 flasks, maintained and grown in 25ml of
MEM (containing 10% FCS, Na+ Pyruvate, Penicillin and Streptomycin, and L-Glutamate), and typically passaged at 1:3, every three to four days.
Due to poor timing last week, I had to split the cells again within two days of their previous passage, and whilst they grew well enough to be split again on Tuesday this week, are now growing poorly. I'm going to leave them over the weekend, and replace their media, however I would like to inquire as to whether this is normal? Will they gradually break back into the cycle of cell division, and what would the implications of this be with regards to transfection efficiency?
(I will be transfecting the cells over the course of a day with PGL3 vector + insert for a Luciferase assay).
Thanks in advance
Chris
Hi, actually, splitting the cells should be based on the confluence of the cells. If they are not confluent and you split them, it will take more to have confluent flask, which might change the cells behavior. Those cells need cell-cell connection to be " Happy" cells.
If that's the case they should be fine, however I have been told they're not supposed to be toughing/clumped with each other, and require a vast amount of space to be happy :S
I do not know why they should not ! As far as I know cell-cell connection would be better for this cells.
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