Objective: Adult stem cells such as mesenchymal stem cells (MSCs) are multipotent. Because of their low immunogenicity MSCs significantly can be used in cell therapy, regenerative medicine and clinical applications.MSCs have been used in the transplant of many organs such as liver, kidneys and heart and also are able to treat many disorders. However, isolation of MSCs from adult tissues such as bone marrow and adipose tissues requires invasive procedures and the availability of an appropriate donor. The amount of MSCs that can be gained from a single donor is limited and the capability of these cells for long-term proliferation is quite poor. ESCs can differentiate into MSCs in certain culture conditions. ESC–derived MSCs could provide unlimited MSCs source for several clinical treatments for instance, mesodermal tissues repair and even for improvement of human ESC-derived HSCs engraftment. Moreover, in a study while treated with IFN-g, ESC–derived MSCs expressed lower HLA-DR than BM-derived MSCs.Materials and Methods: In this research, we derived hMSCs from human ESCs. hESCs were cultured on MEFs that were inactivated by mitomycin C, after that hESCs were passaged mechanically using glass beads and formed embryoid bodies in petridishes (EBs), EBs were then spread into well plates using different medias (hESCs media, DMEM with 10% FBS, DMEM with 20% FBS), later cells were passaged into flasks and finally MSCs were characterized by immunolocalisation using confocal microscopy.Results: As a result, the ESC–derived MSCs adhered to the flask and had a spindle-shaped that resembled BMderived MSCs morphology. Also ESC–derived MSCs were positive for CD44, CD105, CD29 markers the same as BM-derived MSCs. Furthermore MSCs were negative for CD31 and CD45 markers as expected.Conclusion: Our ESC–derived MSCs had two of the three criteria to be defined as MSCs; they adhered to plastic in standard culture conditions. Also MSCs expressed specific cell surface antigens and lack certain other antigens. However the MSCs should in addition have the ability to differentiate into osteocytes, adipocytes and chondrocytes that needs further experiments to be done.In conclusion, large amount of MSCs can be derived from clinically compliant hESCs. However, additional in vivo studies are required to direct differentiate MSCs into specific tissue types for clinical use. Also gain more knowledge of how MSCs reduce immune responses after transplantation is necessary.

Objective: There is currently no optimal treatment for glioblastoma multiforme; the most common malignant brain tumor in adults and patient typical survive < 1 year. This poor outcome relates at least in part to the inability to deliver therapeutic agents to the tumor. Recent evidence suggests that stem cells may inhibit proliferation of malignant cells. Human umbilical cord mesenchymal stem cells can be easily obtained and proliferate rapidly in culture. hUCMs are immunologically compatible and amenable to stable transfection. The aim of this study was to determine whether hUCMs can effectively inhibit expansion of human gliomas in a coculture system. Materials and Methods: hUCMs harvested from human umbilical cord were propagated in laboratory. hUCMs and U87 cells were plated at a density of 5×104 in 50 ml drop of the basic culture medium, 1 cm apart from each other. The cells were given one day to attach to the substratum. After one day, basic culture medium containing DMEM / F12 supplemented with 10% FBS, 100U/ml penicillin- streptomycin was added. Cultures were maintained at 37°C in a humidified atmosphere containing 5% CO2. The invasion of U87 cells in various directions was measured by software (analysis@) in micrometer and the data were compared according to the migration of U87 cells in different directions. Results: Proliferation of U87 cells was less pronounced in the vicinity to the hUCM cells. The U87 cells at the vicinity of hUCMs have migrated 153.96 mm while in three other directions they have migrated 212.25 mm, 264.05 mm and 288.72 mm. Conclusion: hUCMs can inhibit the spread of U87 glioma cells, whether inhibition of the growth of U87 cells by hUCM cells has therapeutic advantages needs to be explored.

Objective: Doxorubicin (DOX) has been one of the most prescribed chemotherapeutic drugs for the treatment of variety malignancies. Unfortunatly, use of doxorubicin is associated with cardiomyocytes apoptosis. The exact mechanism of doxorubicin –induced apoptosis has not fully understood yet. Some evidences demonstrated that doxorubicin induced apoptosis of isolated cardiomyocytes proceeds through activation of nuclear factor-kappaB. In the current study we evaluated effects of doxorubicin on some important genes involved in apoptosis and also Nf-kB activity in H9c2 cardiomyoblast cells.Materials and Methods: Cell viability was determined by MTT assay. Apoptosis was assessed one step Quantitative Real Time RT-PCR was used to evaluate the expression of Bcl-2, Bax, cIAP1, caspase-8 and caspase-9. The DNA-binding capacity of NF-kB (P65 subunit) in nuclear extracts was examined by ELISA method.Results: Doxorubicin induced cytotoxicity in H9c2 cells in a dose dependent manner. Real time RT-PCR analysis shows a significantly reduction (42.7% of control level) of the expression level of Bcl-2 after 22 hours treatment with DOX. Moreover, induction of apoptosis was accompanied by increase in pro-apoptotic Bax level.The Bax/Bcl-2 ratio increased 2.83 ± 0.164 – fold upon treatment with doxorubicin. In contrast to remarkable down- regulation of Bcl-2, we did not show a significant change in the cIAP1 mRNA expression. To determine which apoptotic pathway is activated by doxorubicin; we examined the mRNA expression of caspase-8 and caspase-9. Addition of DOX to H9c2 cells increased significantly mRNA expression of caspase-9 (1.227±0.05- fold of control). However, treatment of H9c2 cells with DOX had no effect on the expression of caspase-8.Also, we observed that NF-kB activity in H9c2 cells was sharply increased (6.86-fold) by incubation in the presence of 3mM doxorubicin after 22 hours.Conclusion: Previous studies have shown that depending on the cell models, apoptosis from DOX can follow different pathways. Our findings demonstrated that apoptosis induced by doxorubicin occurred through intrinsic pathway rather than by extrinsic pathway.Also, DOX sharply increased NF-kB activation. It was indicated that NF- kappaB induces multiple factors to regulate apoptosis at many steps along the cell death cascade including cIAP1 and 2, caspase-8 in various cellular model system. Based on these results, it is possible that the increase in nuclear translocation of the transcription factor, NF- kB (p65 subunit), may contribute in inhibition down regulation of antiapoptotic cIAP 1, caspase-8 and consequently, inhibition of extrinsic pathway of apoptosis in H9c2 cells.

Objective: Multipotent stem cells have been detected in some tissues in the adult, participating in normal replacement and repair. There are some evidences to suggest that adipose stromal cells (ADSCs) not only differentiated into mesodermal cells, but also accept the fate of endodermal and ectodermal cell types. ADSCs can be expanded rapidly in vitro and can be obtained by a less invasive method. In this study, we attempted to evaluation the protocol for the induction of ADSCs into neurons.Materials and Methods: ADSCs was isolated and characterized a population of human primate adipose tissue stromal cells containing multipotent progenitor cells. These cells were induced to differentiate into neural-like cells and analyzed for expression of the neural specific markers. Differentiated ADSCs were assessed for Nestin, MAP2 and GFAP expression by immunocytochemistry and semi quantitative RT-PCR techniques. Moreover, MTT assay was applied to detect cell viability.Results: Immunocytochemical analysis demonstrated that ADSCs had large expression of the neural-specific markers following differentiation. Additionally, soon after differentiation, this protocol showed the highest percentage of mature neural-like cells, however with passing time, the neural like state was transient and reversible. MTT assay showed that induction agents led to more cell death.Conclusion: Analysis showed that neural-like cell differentiation potential of ADSCs in chemical induction was rapidly but transitory.

Objective: Modulating the expression of cell adhesion molecules on hematopoietic stem cells (HSCs) of cord blood (CB) may help to improve homing process of CB stem cells.Microenvironment and released mediators are major factors in the process of engraftment. Based on evidence concerning the existence of substance P (SP) receptor on cord blood CD34+cells, we aimed to explore effects of SP on CD44 expression on CB-HSC.Materials and Methods: CD34+cells purified from CB, were cultured in a serum-free liquid culture system. Different concentrations of SP were used in combination with cytokine cocktail of SCF, FL, TPO, IL3 and IL6. Control groups were treated with cytokine cocktail. CD44 expression was enumerated by flowcytometry.Results: Our results show significant increase in frequency of SP treated cells at 10-7M and 10-11 M following 7 days cultivation as compared to control group. Additionally median flowcytometric intensity of CD44 at corresponding culture period and SP concentrations were significantly increased compare to freshly purified CD34+cells at day 0.Conclusion: Our findings suggest that SP neuropeptides could act as a novel supplement for cytokine cocktails to maintain or increased CD44 expression which could be beneficial in homing of cells in transplantation.