CHO suspension cells - adaptation to serum free suspension, and transfection using electroporation (Oct/02/2006 )

Hi evryone!
I have tried adapting CHO K1 cell line to serum free, suspension medium in CHO SFM II, but I have no idea whether my cells are now adapted or not!!!! the cells now are in suspension, but I am not sure how they should look (appearance) in suspension conditions!?? they look quite shrunken, although i guess they are not dead!!!
Has anyone worked with this system ??? any insights are welcome!!!!!
thanks
monica

---

how long did u keep your cell line in suspension?

---

i was adapting them in serum free suspension medium for about 4 weeks, replacing the hams f 12 medium slowly with cho-sfm medium.. and then after that they were in this state for about 1 week and then i froze them down.. now again i have thawed an aliquot and want to expand them, n do a transfection , n grow using spinner flasks..

---

what about passage times...?

why don't u try to start culturing your cell line with low serum concentration and then dilute it till u end with free medium...
check cell viability by plating a small cell volume ..

---