Primary Culture Of Human Greater Omentum Preadipocytes And Preliminary Observation Of Expression Of Adipocytokines

Objective:To establish a primary culture method of human greater omentum preadipocytes, to observe effect of TNF-αon the expression of C-reactive protein, adiponectin and NF-κB in human omental adipocytes.Methods:Omental adipose tissue were obtained from research objects under aseptic condition, adipose tissue was separated and digested immediately and obtained cell suspension into culture flask was cultured in cell incubator. After inducing differentiation, preadipocytes and differentiated adipocytes were observed by oil red O stain and RT-PCR. Mature adipocytes was treated by TNF-αin different concentrations. The expression of C-reactive protein, adiponectin and NF-κB in human omental adipocytes was detected by RT-PCR.Results:1. Human preadipocytes were primary cultured and inducing differentiated successfully.2. The expression of C-reactive protein mRNA detected by RT-PCR was existent. 3. With TNF-αintervention, the differences between treated group and 0ng/ml group had statistical significance (P<0.05). At the same time, the differences between 20ng/ml and 5ng/ml had significant statistical significance (P<0.01), 20ng/ml and 10ng/ml group, 10ng/ml and 5ng/ml group (P<0.05). At the same concentration, the differences between 48h and 12h group had significant statistical significance (P<0.01), other time groups (P<0.05). At 12h, the differences of CRP, NF-κB mRNA between 10ng/ml and 5ng/ml group had not statistically significant (P>0.05). At 5ng/ml, the differences of adiponectin mRNA between 24h and12h group, 48h and 24h group had not statistically significant (P>0.05), also NF-κB mRNA between 24h and 12h group (P>0.05). The expression level of CRP mRNA was negatively correlated with adiponectin mRNA ( r = -0.9921, P<0.001). The expression level of CRP mRNA was positively correlated with NF-κB mRNA (r = 0.9892, P<0.001). The expression level of adiponectin mRNA was negative correlated NF-κB mRNA (r = -0.9980, P<0.001).Conclusions:Primary culture method of human greater omentum preadipocytes was established successfully and mature adipocytes wered obtained. In human omental adipocytes, the expression level of CRP and NF-κB mRNA was increased with TNF-αconcentration and time-dependent, and the expression level of adiponectin mRNA was decreased.