Abstract: AIM: Mouse embryonic stem cells (mESC) are routinely cultured in the medium with high concentration of glucose (25 mmol/L).However, it was found that chronic high glucose culture promoted the apoptosis of stem cells, inhibited the differentiation efficiency and decreased the response of differentiated beta cells on glucose stimulation.The present study was designed to find suitable lower concentration of glucose to optimize the ESC culture medium so as to improve the ESC growth and differentiation.METHODS: The routinely used high glucose (25 mmol/L) medium was changed to 5, 10, 15, 25 mmol/L respectively at 4 or 12 hours after mESC passage.The colony formation by alkaline phosphatase (AP) staining, cell counting by Trypan blue, cell viability by 3-[ 4, 5-Dimethyl-2-thiazol]-2, 5-diphenyl-2H-tetrazolium bromide (MTT) method were performed.RESULTS: mESCs were changed culturing with different glucose concentrations at 12 hours after passage:(1) There was no significant difference on colony numbers between 5, 10, 15 mmol/L groups and 25 mmol/L group, and AP staining was strongly positive in all groups.(2) There was no significant difference of cell numbers and cell viability at 15 mmol/L group compared with 25 mmol/L group.CONCLUSION: 15 mmol/L glucose, which was used at 12 hours after mESC passage, did not affect the cell viability, pluripotency and undifferentiated state.

Key words: embryonic stem cells, glucose, concentration