Difficulty of thawing H9c2 - (Feb/19/2011 )

Dear All,

I have been facing with difficulty of thawing H9c2 cell line, which was kept in -80C freezer for 6 months.

After 24 hour of thawing, the cells were not attached to the flask, most of them was floating. Just few of them attached to the flask but not looked healthy at all, still rounded shape.

Could anyone give me any suggestions?
Thank you very much.
Sarawut

It sounds like the cells are not viable. The first question is whether the cells were viable when frozen? The next issue is in what solution they were frozen? The final point is how does the rate of thawing affect the cells?

If I were you, I will keep the cell in the same culture medium and see more cells attach or not,
meanwhile I will thaw another vial.

I think usually it is not a good idea to keep cells in -80 for a long term,
it should be store N2 tank,

if you only have those cells (all in -80, and cannot get new), then thaw another, check viable cells number and culture in small well coated with gelatin.

Thank you very much for your valuable suggestions.

Cells were kept in 90%FBS+10%DMSO and quicly thawing in 37C waterbath, right after taking from freezer.

I do agree with you that freezing the cell at -80C freezer is not a good idea.

I will try to thaw another vial and let's see what happend.

Thank you very much indeed.

Sarawut