RESUMO
Objective: The use of stem cells, particularly mesenchymal stem cells [MSCs], with genes and various growth factors as treatments for myocardial infarction and various other diseases is highly regarded. However these cells meet with inflammation and a hypoxic environment in the target tissue. Hence, treatment with factors that increase the resistance of these stem cells is of importance. Stem cells also can be used as carriers for gene therapy. The aim of the present research is to produce VEGF expressing MSCs. We investigate the effect of stromal derived factor 1 on MSC survival in order to use these cells in a future rat myocardial infarction model
Methods: MSCs were purified from young male rats by aspirating the cavity of femurs and tibias. After characterization, MSCs were transduced with VEGF using lipofectamine. Expression and function of VEGF was confirmed. Next, we treated MSCs with SDF1alpha at various time points. The effect of this chemokine was investigated using the LDH assay and by viable cell counts
Results: The experiments confirmed the production and function of VEGF by MSCs. The LDH levels decreased significantly in SDF1alpha treated MSCs. Cell viability increased significantly in the presence of this chemokine
Conclusion: Treatment of MSCs with the SDF1alpha chemokine has increased the survival of these cells. These MSCs are proper candidates for increasing angiogenesis and for further analysis in a rat model of myocardial infarction
RESUMO
Cancer/testis genes [CT-genes] are a family gene which only express in normal testis tissue; some of them are randomly expressed in some types of cancers. The aim of this study was to evaluate the frequency of expression of CT-genes in the patients with breast cancer. Thirty two breast cancer tissue samples were prepared. Expression of NY-ESO-1 1a, NY-ESO-1 1b, SCP1, SSX-2 and MAGE-3 genes, as well as GAPDH [internal control], were studied by multiplex RT-PCR method. Three [9%] of 32 tumor samples expressed mRNA of NY-ESO-1 1a, while six [19%] of 32 tumor samples expressed mRNA of NY-ESO-1 1b. Seven [22%] and two [6%] of 32 tumor samples expressed mRNA of SCP1 and mRNA of MAGE3, respectively. Overall, Thirteen [41%] samples expressed one of the studied genes. NY-ESO-1 and SCP1 genes had the highest frequency of expression of mRNA. It is suggested that more number of breast cancer tumor samples should be examined to evaluate expression of CT-genes. SCP1 and NY-ESO-1 proteins may promote future breast cancer immunotherapy
RESUMO
In recent years, recombinant monoclonal antibodies and their derivatives have emerged as important targeted therapy agents. Monoclonal antibodies are ex-tremely difficult to produce. So, the cost of production is very high and many people cannot afford these drugs. In this regard, choosing inexpensive and easy ways to manipulate production systems such as bacterial hosts to reduce the cost of manufacturing these critical components are considered as vital step for developmental issues in recombinant expression systems. We, therefore, at-tempted to generate a polycistronic construct of anti HER-2 F[ab']2 fragment antibody for insertion in an expression bacterial plasmid. Also some modifica-tions were made in the hinge region to express antibody F[ab']2 fragment in its authentic form preventing from multiple varieties of disulfide bond formation. Finally, synthesized construct was cloned in pET-32 Ek/LIC vector without using restriction enzyme digestion or ligation reactions. The results of this study showed that modified F[ab']2 fragment was simply and successfully inserted in Escherichia coli [E.coli] using the Ligation Independent Cloning technology