Characterization of Adipocytes Differentiated from Primary Human Dermal Fibroblasts of Normal and Diabetic Patients

Background and Aim: The sharp rise in the incidence of diabetes andobesity has reached an epidemic state worldwide. Obesity is one of therisk factors for onset of type 2 diabetes, it also can affect the cardiovascularhealth leading to the onset of cardiometabolic disease. The expansionof white adipose tissue in obese individuals can affect both lipid andglucose metabolism. Diabetic patients often suffer from increase adiposetissue that is defective in glucose metabolism. Therefore, it is essentialto examine the cellular and molecular changes in adipocyte of humandiabetic patients. Since obtaining human adipocytes from patients arechallenging, we aimed to establish protocols for the differentiationof human skin fibroblast cells from normal and diabetic patients toadipocytes. Our main objective was to characterize these adipocytesand determine whether they maintain the diabetic phenotypes afterdifferentiation.Methods: In our current study, we differentiated primary human skinfibroblast cells from normal and diabetic patients into adipocyte after10 days culture in differentiation media. Oil Red O staining was utilizedto assess differentiation to adipocytes. Real-time PCR was used to assesschanges in gene expression.Results: Oil red O staining showed a significant accumulation of lipiddroplets in the differentiated cells. Real-time PCR of non-differentiatedskin fibroblast showed a significantly higher mRNA level of Adiponectin,PPAR gamma and CEPB beta of diabetic patients compared to normal.After differentiation the level of Adiponectin mRNA in diabetic cellsfurther increased however, there was no difference in CEBP beta mRNAlevel in diabetic as compared to normal human differentiated adipocytes.Furthermore, we observed a significant increase In FABP mRNA levelin diabetic patient accompanied by a significant decrease in GLUT-4,insulin receptor and insulin receptor substrate 1 mRNA.Conclusions: Our data illustrates that adipocytes differentiated from skinfibroblasts of diabetic patients maintain the characteristics of the diseasestate. These adipocytes could be used for further biochemical studies.

کلیدواژه ها:

Dermal fibroblast differentiation ، Adipocytes ، Diabetes ، Gene expression