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Background: Human embryonic stem cells (hESCs) are capable of self-renewal and large-scale expansion. They also have the capacity to differentiate into a variety of cell types including liver, cardiac and neuron cells. However, it is not yet clear whether hESCs can differentiate to hemangioblasts under in-vitro conditions. Hemangioblasts are bipotential progenitors that can generate hematopoietic lineages and endothelial cells. The aim of this study was to identify the potential of human Royan H5 embryonic stem cells in differentiating into hemangioblast cells. Methods: HESCs were cultured at suspension system in DMEM/F12 supplemented with bFGF. 7-day old cells differentiated into blast cells under defined condition consisting of hematopoietic cytokines including BMP4, VEGF, etc. Blast cell markers kinase insert domain receptor (KDR), CD31, and CD34 were evaluated by flow cytometry and blast gene expressions (TAL-1, Runx-1 and CD34) were detected by qRT-PCR. Clonogenic assays were performed in semisolid medium by colony forming unit-assays. Results: The hESCs (Royan H5) had the capacity of differentiating into hemangioblast cells. We could detect colonies that expressed 79%±12.5 KDR+, 5.6%±2.8 CD31+-CD34+ and 6%±2.12 KDR+-CD31+ on day 8 in the hESCs. Up-regulation of TAL-1, Runx-1 and CD34 occurred during hemangioblast commitment (P≤0.05 and P≤0.01, respectively). Moreover, hemangioblast cells generated mixed-type and endothelial-like colonies in semi-solid media. Conclusion: Our results showed that hESCs (Royan H5) were able to differentiate into hemangioblasts under in-vitro conditions. The hemangioblasts had the potential to generate two non-adherent (Mixed-type) and adherent (endothelial-like) cell populations.

"n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1":*{behavior:url(#ieooui) } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Systemic scleroderma (SSc) is a generalized connective tissue disorder of unknown origin which most notably is characterized by skin thickening and organ damage. Endothelin-1 (ET-1) antibody plays a role in skin fibrosis. The aim of this study was to determine the prevalence and correlation of different manifestations of SSc with ET-1 plasma levels."n"nMethods: This cross-sectional analytical study was conducted on 95 patients (91 women and four men) with scleroderma in 2006. The patients had been referred to the Rheumatology Clinic of Shariati Hospital in Tehran, Iran. The demographic data and signs and symptoms were entered in a questionnaire and endothelin-1 concentrations were measured. "n"nResults: The mean age of the patients was 38±12.29 years. Diffuse cutaneous SSc (dcSSc) was diagnosed in 52 and limited cutaneous SSc (lcSSc) in 43 patients. Raynaud's phenomenon (91%) was the most common manifestation in the patients. The relationship between the resorption of terminal phalanges due to fibrosis with the plasma concentration of Endothelin-1 was statistically significant (p=0.001). Pitting ulcers had significant relationships with endothelin-1 concentrations too (p

Background and Objectives The production of a sufficient number of pancreatic endocrine cells that function similarly to primary islets is one of the methods in treatment of diabetes. The CD 133+ cells derived from cord blood (UCB-CD133) express some embryo specific markers such as SSEA-4 and OCT4 and so it can be a candidate for differentiation into insulin secreting cells. Therefore, here we attempt to determine the differentiation potential of UCB-CD133+ cells into insulin secreting cells in vitro. Materials and Methods UCB-CD133+ cells were isolated by MACS and differentiated into insulin secreting cells using the medium containing DMEM/F12, bFGF, Nicotinamid, and supplement B27 N2 for 9 days. The expression of insulin and c-peptide protein was detected by immonocytochemistry. The expression of insulin, Nkx6.1, Pdxl, and Glocagon genes was analyzed by RT-PCR; ELISA was performed to analysis the function of differentiated cells in different glucose concentrations (5 and 27 mM). Results Our results determined that UCB-CD133+ cells expressed insulin and c-peptide at protein level after 9 days of culture. However, the insulin, Nkx6.1, Pdxl, and Glocagon genes were not detected in differentiated cells and they could not respond to different concentrations of glucose. Conclusions We suggested that UCB-CD133+ cells can differentiate into insulin secreting cells in vitro; however, they are not functional and need to receive more signals in vivo. [ABSTRACT FROM AUTHOR]

Objective: The transmembrane receptor family of Tolllike receptors (TLRs) is expressed in immune cells and cardiac muscle. To date, several TLRs (numbered 1-11) have been identified in humans. TLRs may play a role in initiating early inflammatory and functional responses to danger signals arising from ischemia-reperfusion and inflammatory stimuli. Human embryonic stem cells (hESCs) have the potential to provide an unlimited source of cardiomyocytes, which are invaluable resources for drug or toxicology screening, medical research, and cell therapy. Materials and Methods: Royan H5 and Royan H6 hESC lines were used in this study. Suspension culture of hESCs was performed according to a recently published protocol. Differentiation of the cells into cardiomyocytes in suspension was performed according to the Laflamme et al. protocol with some modifications. In this study, expression of TLR2, TLR3, TLR4, TLR5 and TLR9 was evaluated by RT-PCR and Q-PCR during cardiomyocyte differentiation on day 8, 14 and 25 (mature cardiomyocyte). Results: According to Q-PCR data, TLR9 expression has increased and TLR5 expression has decreased in linear pattern during cardiomyocyte differentiation. Expression pattern of TLR2 and TLR4 was same in sigmoid shape. In both of them, expression level was lowest in mature cardiomyosyte. Also, about TLR3 expression, our data was shown sigmoid shape but the lowest expression related to 8th day after differentiation. Conclusion: The goal of this study was to investigate the ability TLR expression in cardiomyocyte differentiation from human embryonic stem cells. According to results, all of TLRs in this study have been expressed in cardiomyocyte in variable level. TLRs expression related to inflammatory responses, ischemia and contractility. On the other hand, myocardial infarction and all of myocardial ischemia diseases have inflammatory reasons. Hence, evaluation of TLRs expression during cardiomyocyte differentiation is indispensable in cardiac cell based therapy. [ABSTRACT FROM AUTHOR]

Regenerative therapies and cell replacement have great perspectives in the treatment of degenerative disorders in which the function or structure of the affected tissues or organs are deteriorated over time. Stem cells, both embryonic and adult, are evaluated as the potential substrates of cell therapy for tissue regeneration in such diseases. In fact, stem cells can be considered revolutionary in the field of medicine. However before using the cell therapy routinely in clinics their potentials have to be studied in clinical trials if there are a reliable data in animal models for safety and effectiveness. The filed of stem cell therapy, had a relatively strong start in Royan institute, benefiting from institute scientific infrastructure. The first clinical trials, using autologous bone marrow derived stem cell, was conducted in patients with myocardial infarction at 2006 and it followed with cell therapy trials in the kidney, vascular, liver, skin, and eye; bone diseases; and neurological disorders as well as progressively proceeding to the final phases of safety and efficacy. Now, there are more than 30 registered clinical trials by Royan institute in clinicaltrials.gov site. Clinical studies in Royan are focused mainly on the use of bone marrow and tissue specific stem cells and there is no any clinical trial using embryonic or fetal stem cell. This presentation reviews the current clinical outcomes about cell therapy in heart, bone and joint and skin disorders which are conducted in Royan institute. [ABSTRACT FROM AUTHOR]

Objective: Scleroderma -one of the important autoimmune diseases- leads to death of endothelial cells as one of the early events of this disease. The lack of repair after the loss of endothelial cells is observed in these patients and although it is a worldwide disease, its cause has remained unknown. Because of physiological differences between human and other species, a proper scleroderma animal model which represent all aspects of the disease has not been generated yet. So, making a patient-based system to mimic a developmental defect and evaluating its probable repair mechanism is considered as a necessity. In this study it was tried to generate patients' specific induced pluripotent stem (iPS) cells and characterize them. Materials and Methods: Patient's fibroblast cells -as an available and unlimited source of disease study in vitrowere isolated, cultured and transduction with retroviral vector containing Oct 3/4, Sox2, Klf4 and c-Myc. For characterization of generated iPS cells, their expression of endogenous pluripotent and silencing of retroviral genes were evaluated by RT-PCR technique and the existence of pluripotency proteins were assessed by immunostaining. Spontaneous differentiation into endothelial cells was done by embroid body formation and the expression of specific endothelial cell markers were examined by flowcytometery technique. Results: RT-PCR analysis and immunostaining study of generated cells compared to human embryonic stem cells (royan H5) results showed that reprogrammed cells express specific markers similar to embryonic stem cells. EB formation results And also, spontaneous differentiadifferentiation into endothelial cells using EB formation showed that our scleroderma derived pluripotent cells have differentiation potential like embryonic stem cells. Conclusion: Our study showed that scleroderma specific iPS cells can be indicated as an unlimited and available source for assessing molecular defects in vitro and finding regenerative methods to repair endothelial cells in scleroderma patients [ABSTRACT FROM AUTHOR]

Objective: Increasing rate of morbidity and mortality by heart diseases all over the world attracted scientists' attention to this important medical case. In recent years regenerative medicine has been introduced as an alternative tool for heart repair; and cardiac stem cells (CSCs), precursor cells resident in heart tissue have been investigated as novel sources of heart cell therapy. Such patient derived stem cells need to be isolated from small heart biopsies and expanded in culture to reach the quantity which is required for cell therapy. Although various aspects of adult stem cell transplantation have been well studied, the effects of extensive sub-culturing on the characteristics and phenotype of these cells have remained elusive. Materials and Methods: Human CSCs and skeletal muscle stem cells (satellite cells) were isolated from heart and intercostals biopsies of patients with congenital diseases during routine heart surgeries with informed consents. Isolated cells were passaged until their morphologies began to change and they were sampled at 4 passages -passage 6, 9, 12 and 15. Cell surface markers' changes were analyzed using flowcytometery system and whole genome transcriptom alteration was assayed using Illumina chip microarray analysis. Results: Doubling time measurement results showed that subsequent passages caused an increase in essential time for cell number duplication and flowcytometry analysis indicated the different cell surface markers at different passages. Whole genome analysis showed gene expression alteration by increasing the passage number (ANOVA p value⩽0.05, fold change⩾2). The most important gene ontology which significantly altered during passages was cell cycle. Conclusion: Our study showed that subsequent passages cause significant changes in both phenotypic and genome expression characteristics of CSCs and satellite cells. As both CSCs and satellite cells are considered as stem cell therapy sources, it's important to decrease any cell modification in culture. [ABSTRACT FROM AUTHOR]

Objective: Acute kidney injury (AKI) is a potentially devastating disease in clinical medicine. However, no specific therapy improves the rate or effectiveness of the repair process after AKI. Bone marrow-derived mesenchymal stem cells (BM-MSCs) have been proven to be benefit to the renal repair after AKI in different experimental models of rodent models, but the consequence of these results to large animals and eventually, to humans remains unknown. Thus, the aim of this study was to assessment the effect of autologous rhesus Macaque monkey BM-MSCs transplantation in cisplatin-induced AKI. Materials and Methods: BM-MSCs were characterized for their growth characteristics, differentiation potential, immounophenotypic properties and chromosome content. According to design procedure, monkeys were divided into control, vehicle and cell transplantation (Cell Tx) groups and exposed to cisplatin 5mg/kg as intravenous for induce of AKI. Control animals were not treated with anything but cell Tx and vehicle animals were treated with intrarenal arterial injection of autologous BM-MSCs and normal saline, respectively. For cell tracking with magnetic resonance imaging (MRI), BM-MSCs labelled with nanoparticles superparamagnetic iron oxide (SPIO). Results: Labelled BM-MSCs were found in both glomeruli and tubules. Transplantation of 5×106 cell/kg ameliorated renal function during first week as reflected by significantly lower serum creatinine and urea values and higher urine creatinine and urea clearance without hyponatremia, hyperkalemia, proteinuria and polyuria to 3 months in compare with vehicle and control groups. BM-MSCs markedly accelerated present of Foxp3+ regulatory T (Treg) cells in response to cisplatin-induced damage, as revealed by higher numbers of Foxp3-positive cells within the tubuli with respect to cisplatintreated monkeys in control and vehicle groups. However, BM-MSCs did not repair kidney parenchyma and as partially to modulate ultrastructure of kidneys. Conclusion: These data demonstrate that BM-MSCs in this unique cisplatin-induced AKI large-animal model, did exhibit reparative and protective properties [ABSTRACT FROM AUTHOR]