AIM: To filter glucose concentration and culture time of Wistar rat retinal vascular endothelial cells(RVEC)in high glucose model.

METHODS: Primary culture of RVEC of Wistar rats with 1-3d of birth,Ⅷ factor antibody immunohistochemistry to identify the cells; Establishment 0mmol/L glucose as a control group(group C), at different concentrations of glucose intervention in turn divided into 10mmol/L(group A), 15mmol/L(group B), 25mmol/L(group D), 35mmol/L(group F), MTT assay was used to detect cell OD values of each group and each group was detected by flow cytometry apoptosis rate were at three time points: 24,48,72h。

RESULTS: RVEC purity was 94%. Effect of different concentrations of glucose in Wistar rat RVEC by MTT assay: after 24h culture, OD value of experimental group decreased, but the difference was not statistically significant(F=0.42, P=0.7411); after 48h culture, OD value of the difference between the groups was statistically significant(F=45.2, P=0.000), pairwise comparison between the five groups, except among groups A, B, C group no difference was not statistically significant external(Q_48AB=44.0427, Q_48AC=36.4701, Q_48BC=27.7953, all P> 0.05), the remaining pairwise comparisons were statistically significant difference between groups(Q_48CD=11.0715, Q_48AD=14.0794, Q_48AF=12.2964, Q_48BD=23.4698, Q_48BF=12.7016, Q_48DF=10.2013, all P<0.05; Q_48CF=6.4701, P<0.01). After 72h culture, OD value of each group compared to the difference was statistically significant(F=37.46, P=0.000), pairwise comparisons among five groups, the same A, B, C no difference among the three groups was not statistically significant(Q_72AB=27.7338, Q_72AC=25.0054, Q_72BC=33.3797, all P>0.05), the rest of the group differences were statistically significant(Q_72AD=13.4793, Q_72BD=12.7546, both P<0.05; Q_72CD=7.3743, Q_72CF=8.3465, Q_72AF=4.7455, Q_72BF=3.9471, Q_72DF=3.2649, all P<0.01). Effect of different concentrations of glucose on RVEC apoptosis rate: after 24h culture, Differences between groups was not statistically significant(P>0.05). After 48h culture, pairwise comparison among five groups, Q_48AB=31.1704, Q_48AC=33.5947, Q_48BC=29.3968, Q_48AD=30.4097, Q_48BD=28.8164, both P>0.05; Q_48CD=12.5032, Q_48AF=11.7531, Q_48BF=14.1076, Q_48DF=13.9372, both P<0.05; Q_48CF=7.0953, P<0.01. After 72h culture, pairwise comparison among five groups, Q_72AB=26.3392, Q_72AC=24.9142, Q_72BC=30.2976, both P>0.05, the rest of the group differences were statistically significant(where Q_72AD=14.1983, Q_72BD=12.9356, both P<0.05; Q_72CD=6.8752, Q_72CF=7.6745, Q_72AF=5.0545, Q_72BF=4.0741, Q_72DF=3.8876, both P<0.01).

CONCLUSION: Best glucose concentration and the time is 25 mmol/L and 48h, for RVEC high sugar model.