I am currently preforming ceiling culture and so far have had no major problems, though my supervisors are arguing they could be stromal contamination. I will repeat again, this time marking cells before starting cultures.
Are you spinning your samples enough? I found that removing the lower layer containing the stromal portion and keeping the adipocytes in the same tube and washing and spinning in the same tube everytime reduces loss of adipocytes.
I know you posted this in May, I hope you have had some success by now.
do you use lipoaspirate or solid tissue (e.g. omental fat tissue) as starting sample? do you know if lipoaspirate has been used to obtain mature adipocytes culture?
Are you still working with ceiling culture of mature adipocytes? I am writing a grant proposal that would involve using this method and I need a patner/supervisor to team up with and a laboratory visit. Thank you.