I've been working with HBE cells over a year now and it hasn't really gotten any easier. I was wondering if any of you have worked with these cells. After spending hours with ATCC over the phone...I'm still a little confused.
I understand how to split the cells (especially how to neutralize them) but now I wonder about my medium. K-SFM protocol says to add the supplements when needed. However, i find this impractical considering the very small volumes of the supplements. Any ideas? How do you do it? (Dilute the EGF in water I guess?)Also it states to keep the K-SFM in the dark especially once it is supplemented. Should I cover it with foil? Also, once with the cells...is it still necessary (to keep the medium in darkness)? Any help would be greatly appreciated. The more specifics the better!
P.s. I've attached the official protocols for both HBE and K-SFM.