Dear researchers,
Recently, I have developed a study of natural drugs to alleviate high glucose-induced conditionally immortalised murine podocyte injury. Unfortunately, after nearly one month of experimentation, I still could not successfully induce a high glucose damage model.
I read a lot of literature and tried the methods that so many researchers used. These methods include: serum starvation (means serum-free 24 h) + 10% FBS + different concentrations of high glucose(from 30 mmol to 60 mmol (one gradient per 5 mol)), serum starvation (24 h) + 2% FBS + different concentrations of high glucose(from 30 mmol to 60 mmol (one gradient per 5 mol))，serum starvation (24 h) + 1% FBS + different concentrations of high glucose(from 30 mmol to 60 mmol (one gradient per 5 mol)), serum starvation (24 h) + 0% FBS + different concentrations of high glucose(from 30 mmol to 60 mmol (one gradient per 5 mol)).
10% or 2%FBS+high glucose, the podocytes didn't apotosis or death (podocyte cells growth very nice, it's so crazy), the model group(high glucose) and control group (normal glucose 5.5mmol or 11.1mmol) were no different. 1% or 0%FBS+high glucose, model group podocyte were dead, but the normal group podocyte also dead, no different. The methods to detect the difference between the two groups in this study contained：CCK-8 and JC-1/Hoechst 33342 Fluorescent staining and the time to detect contained:24h, 48h, 72h.
My MPC5 cells cultured under 33°+10U IFN-γ to proliferate and under 37°-IFN to differentiation(14d, then used to experiment). I don't know why I can't induce high glucose damage model. Anyone can help me? Thanks a lot lot lot lot lot.
Xiaotao Zhou, have you solved this problem? Same is happening to me!!