Journal of

Background and Aim: MicroRNAs (miRNA) are small noncoding RNA molecules that transcribed by RNA polymerase II. After biogenesis, these molecules act by incorporation into the RNA-induced silencing complex (RISC). MiRNAs are involved in multiple physiological and pathological processes such as proliferation, differentiation, apoptosis and cancer.
Recently several studies reported down regulation of mir-150 during erythropoesis. Since hemoglobin expression is valuable indicator of erythroid differentiation we evaluated the mir-150 downregulation effect on alpha chain expression by Quantitative RT-PCR.

Materials and Methods: K562cells were grown in RPMI1640 in standard condition. K562 cells were transfected by microRNA 150 Inhibitor using transfection kit .Mir-150 downregulation was confirmed by miRNA Real time PCR, followed by Q-RT-PCR to investigate the alpha chain expression changes.

Results: By relative QRT-PCR the alpha chain expression was increased 10 folds in comparison to untransfected and scramble cells. Furthermore, the differences were statistically significant (P<0.05)

Conclusion: Elevation of alpha chain expression in our study showed that mir-150 downregulation has a crucial role in erythroid differentiation and can introduce as a novel marker in alpha thalassemia. Further researches to find out the detail mechanism and miRNAs genes target could improve our knowledge about miRNAs potential in management of diseases and their applications in gene therapy and regenerative medicine.

Background and Aim: Embryonic stem cells are identified with two unique characteristics. First, they can be maintained and expanded as pure populations of undifferentiated cells, a characteristic which is known as self renewal aspect of embryonic stem cells. Second, these cells can give rise to all body cell types. In the current study, we used a feeder-free condition to differentiate mouse embryonic stem cells into lymphoid lineage by IL-7 and FLT-3 ligand.

Materials and Methods: Mouse embryonic stem cells cultured on mouse embryonic fibroblasts were separated from the feeder layer. Then, embryoid bodies were formed from mouse embryonic stem cells. Following that, differentiation was performed by FLT-3 ligand and IL-7. In order to demonstrate the differentiation into lymphoid lineages, the expression of CD25, CD19 and CD3 was assessed by RT-PCR technique on days 7 and 14.

Results: After 14 days of differentiation into lymphoid lineages by defined factors, RT-PCR results showed the expression of CD25 and CD19 markers.

Conclusion: In all previous studies, mouse embryonic stem cells were differentiated into lymphoid lineage by OP9 stromal feeder cells. In this study, a feeder-free condition was used to differentiate mouse embryonic stem cells into lymphoid lineage. It is hoped that the present study can lead to new insights in cell therapy of lymphoid deficiency disorders.

Background and Aim: Obesity is now considered as one of the main risk factors of certain known diseases such as cardio-vascular diseases, non- insulin-dependent diabetes, and common cancers. Moreover, the increase of white fat tissue is known as a main factor in the obesity process, in terms of physiology and pathology. Therefore, the understanding of adipocytes differentiation processes is crucial.

Materials and Methods: In this study, mesenchymal stem cells (MSCs) were isolated from human bone marrow by ficol-gradient, and then, their surface markers were confirmed by flow cytometry. Osteoblastic and adipocytes differentiation were done by dexamethasone protocol and confirmed by staining. Then qualitative and quantitative expressions of PPARgamma (PPAR-γ) gene as an important transcription factor in the differentiation of fat were studied by RT-PCR and REAL TIME PCR before and after differentiation into adipocytes. For statistical analysis, paired t-test was performed, using pfaffl and graph pad software.

Results: PPAR-gamma gene expression showed a significant increase after differentiation of human bone marrow mesenchymal stem cells into adipocytes (p<0/05).

Conclusion: According to the results, the PPAR-γ gene acts as one of the important factors in the differentiation of MSCs into adipocytes. In brief, the inhibition of this gene's expression to prevent obesity is suggested as an idea for treatment in the future.