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Ghasemi-Mobarakeh L.,Isfahan University of Technology | Prabhakaran M.P.,National University of Singapore | Nematollahi M.,Royan Institute for Biotechnology | Karbalaie K.,Royan Institute for Biotechnology | And 2 more authors.
International Journal of Polymeric Materials and Polymeric Biomaterials | Year: 2014

With recent advances in developmental and stem cell biology, the application of stem cells in tissue engineering has received great attention and designing of suitable scaffolds to support cell growth, differentiation, and functional tissue organization are advancing toward effective tissue regeneration. Regeneration of the infarct myocardium after myocardial infarction (MI), which is caused by the abrupt occlusion of one or more of the coronary arteries in the heart is one of the most demanding aspects in tissue engineering. Embryonic stem cells (ESCs) can differentiate into many cell types and has been considered as a cell source for cardiac regeneration. In this regard, nanofibrous scaffolds received great attention in tissue engineering field due to their similarity in morphology to native extracellular matrix (ECM) and various scaffolds have been studied as cardiac patches over the previous years. In this study poly (ε-caprolactone) (PCL)/gelatin nanofibrous scaffolds were fabricated by electrospinning and embroyonic bodies (EBs) were formed using ESCs seeded on the nanofibrous scaffolds. SEM images revealed cell outgrowth from EBs and the spreading of cells over the nanofibrous scaffolds were observed. Immunocytochemistry results showed the cellular expression of cardiac proteins, namely α-actinin and connexin 43 on the nanofibrous scaffolds indicating the differentiation of EBs to cardiomyocytes. Results of our study showed that PCL/gelatin nanofibrous scaffolds can act as a promising substrate for differentiation of EBs to cardiomyocytes and could be applied for cardiac tissue engineering. © 2014 Taylor and Francis Group, LLC.


Honardoost M.A.,University of Isfahan | Kiani-Esfahani A.,Royan Institute for Biotechnology | Ghaedi K.,University of Isfahan | Ghaedi K.,Royan Institute for Biotechnology | And 4 more authors.
Gene | Year: 2014

Background: Multiple sclerosis is an inflammatory autoimmune disease widely characterized by myelin destruction of CNS. Th-17 cells, have been demonstrated to play a crucial role in pathogenesis of MS. MicroRNAs are a new class of non-coding RNAs that participate in post-transcriptional regulation of gene expression. Previous studies have reported a potential role of various miRNAs in induction of Th-17 differentiation and progress of autoimmune diseases. In recent years, it has been shown that miR-326 and miR-26a involved in progress of Th-17 and MS disease. Objective: To evaluate expression pattern of miR-326 and miR-26a in peripheral blood lymphocytes of relapsing-remitting MS patients during relapsing and remitting phases compared to healthy control subjects. Materials and methods: Forty RR-MS patients of Isfahan population were diagnosed as relapsing (n. = 20) or remitting phase (n. = 20) patients according to clinical manifests and expression level of miR-26a and miR-326 was measured in these groups by quantitative real time PCR method compared to 20 healthy controls. In-silico molecular signaling pathway enrichment analysis was also performed on validated and predicted targets (targetome) of miR-26a by DAVID database to explore possible role of miR-26a in Th17 differentiation. Results: We observed up-regulation of both miR-326 and miR-26a in relapsing phase of multiple sclerosis patients compared with remitting phase (p value. = 0.0001) and healthy controls (p value. = 0.0091). ROC curve analysis confirmed valuable and precise potential of miR-326 to discriminate between relapsing and remitting phases of multiple sclerosis with specificity and sensitivity of 100% at a proposed optimum cutoff point. Furthermore, in-silico molecular signaling pathway enrichment analysis detected TGF-β signaling pathway as one of the most statistically relevant pathway with miR-26a targetome. Conclusion: Our results confirmed potential of miR-326 as a diagnostic biomarker to discriminate between relapsing and remitting phases of multiple sclerosis disease. Similar expression pattern to miR-326 and in-silico molecular enrichment analysis altogether suggest an inducing role of miR-26a in differentiation of pathogenic Th17 cells during pathogenesis of multiple sclerosis by targeting major components of the TGF-β signaling pathway (i.e. SMAD4 and SMAD1) and disarrangement of this signaling pathway. © 2014 Elsevier B.V.


Hosseini S.M.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Biotechnology
Reproductive BioMedicine Online | Year: 2016

In October 2012, the American Society for Reproductive Medicine (ASRM) and, in March 2012, the European Society of Human Reproduction and Embryology (ESHRE), lifted the categorization of oocyte cryopreservation as being "experimental" and endorsed its entrance into the mainstream of assisted reproductive techniques. This change in policy, with the considerable advantages that oocytes offer over embryos for cryopreservation, has increased applications of oocyte cryopreservation in assisted reproduction techniques. A deep understanding of oocyte cryobiology, however, is lagging behind the forces propelling the clinical application of oocyte cryopreservation. We have drawn attention to this shortcoming by initiating a debate on whether a vitrified-warmed oocyte has the same characteristics as its fresh sibling. The answer to this question may explain why the oocyte cryopreservation success rate is as yet far from satisfactory and why cryopreserved oocytes should be treated differently from their fresh siblings. A fresh look at the characteristic features of oocytes after cryopreservation is the main scope of this review as a stimulus to further improvement of oocyte cryopreservation. © 2015.


Esfahani M.H.N.,Royan Institute for Biotechnology | Tavalaee M.,Royan Institute for Reproductive Biomedicine
International Journal of Fertility and Sterility | Year: 2012

Varicocele is considered as one of the main etiologies of male infertility. Along with altered semen parameters, increased DNA fragmentation is believed to play an important role in varicocele-induced infertility. DNA damage may result from intra- or extra-testicular factors. Among these, apoptosis, abnormal chromatin packaging and oxidative stress are the most researched and are addressed in this review. Significant evidence suggests that varicoceles have a harmful effect on testicular function and a varicocelectomy not only prevents progressive decline in testicular function, but also reverses the damage. However, the degree to which varicocele repair improves pregnancy rates and the success of assisted reproductive technology (ART) remains controversial. Therefore, the role of varicocele repair on DNA fragmentation is also discussed.


Khazaie Y.,Royan Institute for Biotechnology | Nasr Esfahani M.H.,Royan Institute for Biotechnology | Nasr Esfahani M.H.,Isfahan Fertility and Infertility Center
International Journal of Fertility and Sterility | Year: 2014

MicroRNAs (miRNAs) are small non-coding single stranded RNA molecules that are physiologically produced in eukaryotic cells to regulate or mostly down-regulate genes by pairing with their complementary base-sequence in related mRNA molecules in the cytoplasm. It has been reported that other than its function in many physiological cell processes, dysregulation of miRNAs plays a role in the development of many diseases. In this short review, the association between miRNAs and some male reproductive disorders is surveyed. Male factor Infertility is a devastating problem from which a notable percentage of couples suffer. However, the molecular mechanism of many infertility disorders has not been clearly elucidated. Since miRNAs have an important role in numerous biological cell processes and cellular dysfunctions, it is of interest to review the related literature on the role of miRNAs in the male reproductive organs. Aberrant expression of specific miRNAs is associated with certain male reproductive dysfunctions. For this reason, assessment of expression of such miRNAs may serve as a suitable molecular biomarker for diagnosis of those male infertility disorders. The presence of a single nucleotide polymorphism (SNP) at the miRNAs' binding site in its targeted mRNA has been reported to have an association with idiopathic male infertility. Also, a relation with male infertility has been shown with SNP in the genes of the factors necessary for miRNA biogenesis. Therefore, focusing on the role of miRNAs in male reproductive disorders can further elucidate the molecular mechanisms of male infertility and generate the potential for locating efficient biomarkers and therapeutic agents for these disorders.


Hosseini S.M.,Royan Institute for Biotechnology | Hajian M.,Royan Institute for Biotechnology | Moulavi F.,Royan Institute for Biotechnology | Asgari V.,Royan Institute for Biotechnology | And 2 more authors.
Cellular Reprogramming | Year: 2013

The potential applications of a simplified method of somatic cell nuclear transfer (SCNT) that is improved in both efficiency and throughput is considerable. Technically, a major step of SCNT is to produce large pools of enucleated oocytes (cytoplasts) efficiently, a process that requires considerable micromanipulation skill and expensive equipment. Here, we have developed an efficient and high-throughput method of manual oocyte enucleation using a simple device, a pulled Pasteur pipette, that can be connected to standard zona-free method of embryo reconstitution. Common Pasteur pipettes were pulled on a flame to produce finely drawn pipettes with inner diameters approximately less than half the oocyte diameter (∼50-60 μm), and slightly larger than cytoplasmic protrusion (∼20-30 μm) that was induced after demecolcine treatment of MII-stage oocytes. Oocyte manipulation was performed under a stereomicroscope by either bisecting the oocyte into two approximately equal demioocytes (blind manual enucleation), or by positioning the oocytes so that the cytoplasmic extrusion that contains the MII chromosome mass is removed with the minimum amount of cytoplasm (oriented manual enucleation). The survival rate of the manually enucleated oocytes was 81.4-91.5%, comparable to standard zona-free method of oocyte enucleation (>95%). A total of 80-120 oocytes could be enucleated in 10 min, which was considerably higher than standard zona-free enucleation method. In vitro development rates of cloned embryos derived from manually enucleated cytoplasts with varying cytoplasmic volumes (50%, 95%, and 100%) was comparable, and embryonic developmental rates of the two latter groups were at least as good as standard zona-free method. The manual method of oocyte enucleation described here can be learned and mastered for simple, fast, and cheap production of cloned embryos with comparable efficiency to other available methods. © 2013, Mary Ann Liebert, Inc.


Meamar R.,Islamic Azad University at Najafabad | Nasr-Esfahani M.H.,Royan Institute for Biotechnology | Mousavi S.A.,Isfahan University of Medical Sciences | Basiri K.,Isfahan University of Medical Sciences
Journal of Clinical Neuroscience | Year: 2013

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder of upper and lower motor neurons, characterized by progressive muscular atrophy and weakness which culminates in death within 2-5 years. Despite various hypotheses about the responsible mechanisms, the etiology of ALS remains incompletely understood. However, it has been recently postulated that stem cell therapy could potentially target several mechanisms responsible for the etiology of ALS and other nervous system disorders, and could be regarded as one of the most promising therapeutic strategies for ALS treatment. We present a brief review of different methods of stem cell therapy in ALS patients and discuss the results with different cell types and routes of administration. © 2013 Elsevier Ltd. All rights reserved.


Deemeh M.R.,Royan Institute for Biotechnology | Tavalaee M.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Biotechnology | Nasr-Esfahani M.H.,Isfahan Fertility and Infertility Center
Reproductive Sciences | Year: 2015

Artificial oocyte activation (AOA) has shown to improve fertility in severe male infertility following intracytoplasmic sperm insemination (ICSI). However, the effect of AOA on the health status of children has not been studied. This pilot historical cohort study aims to evaluate physical and mental health of 79 and 89 children from 275 and 406 couples undergoing ICSI-AOA using ionomycin and conventional ICSI, respectively. The outcomes assessed were clinical pregnancy, abortion, type of delivery, and health of children (major birth defect, mental and behavior status). No significant differences were observed between the ICSI-AOA and the ICSI groups for these parameters, and the rate of major birth defects were not significantly different between the 2 groups. In this study, AOA has not imposed a greater risk on physical and mental health of children born through AOA, but for such a solid conclusion, further trails with higher number of cases are required and conclusions drawn are limited to this study. © The Author(s) 2014.


Masaeli E.,Isfahan University of Technology | Masaeli E.,Royan Institute for Biotechnology | Masaeli E.,University of Twente | Morshed M.,Isfahan University of Technology | And 6 more authors.
PLoS ONE | Year: 2013

Tissue engineering techniques using a combination of polymeric scaffolds and cells represent a promising approach for nerve regeneration. We fabricated electrospun scaffolds by blending of Poly (3-hydroxybutyrate) (PHB) and Poly (3-hydroxy butyrate-co-3- hydroxyvalerate) (PHBV) in different compositions in order to investigate their potential for the regeneration of the myelinic membrane. The thermal properties of the nanofibrous blends was analyzed by differential scanning calorimetry (DSC), which indicated that the melting and glass temperatures, and crystallization degree of the blends decreased as the PHBV weight ratio increased. Raman spectroscopy also revealed that the full width at half height of the band centered at 1725 cm-1 can be used to estimate the crystalline degree of the electrospun meshes. Random and aligned nanofibrous scaffolds were also fabricated by electrospinning of PHB and PHBV with or without type I collagen. The influence of blend composition, fiber alignment and collagen incorporation on Schwann cell (SCs) organization and function was investigated. SCs attached and proliferated over all scaffolds formulations up to 14 days. SCs grown on aligned PHB/PHBV/collagen fibers exhibited a bipolar morphology that oriented along the fiber direction, while SCs grown on the randomly oriented fibers had a multipolar morphology. Incorporation of collagen within nanofibers increased SCs proliferation on day 14, GDNF gene expression on day 7 and NGF secretion on day 6. The results of this study demonstrate that aligned PHB/PHBV electrospun nanofibers could find potential use as scaffolds for nerve tissue engineering applications and that the presence of type I collagen in the nanofibers improves cell differentiation. © 2013 Masaeli et al.


Moulavi F.,Royan Institute for Biotechnology | Hosseini S.M.,Royan Institute for Biotechnology | Hajian M.,Royan Institute for Biotechnology | Forouzanfar M.,Royan Institute for Biotechnology | And 5 more authors.
Reproduction | Year: 2013

The effect of technical steps of somatic cell nuclear transfer (SCNT) on different aspects of cloned embryo development was investigated in sheep. In vitro-matured oocytes were enucleated in the presence or absence of zona and reconstituted by three different SCNT techniques: conventional zona-intact (ZI-NT), standard zona-free (ZF-NT) and intracytoplasmic nuclear injection (ICI-NT). Stepwise alterations in nuclear remodeling events and in mRNA abundances, throughput and efficiency of cloned embryo development and cell allocation of the resulted blastocysts were assessed. Early signs of nuclear remodeling were observed as soon as 2 h post-reconstitution (hpr) for fusion-based methods of nuclear transfer (ZI-NT and ZF-NT) but were not observable until 4 hpr with the ICI-NT method. The relative mRNA abundances of HSP90AA1 (HSP90), NPM2 and ATPase genes were not affected by i) presence or absence of zona, ii) oocyte enucleation method and iii) nuclear transfer method. After reconstitution, however, the relative mRNA contents of POU5F1 (OCT4) with the ZI-NTand ZF-NT methods and of PAPOLA (PAP) with ZF-NTwere significantly lower than those for the ICI-NT method. Zona removal doubled the throughput of cloned blastocyst development for the ZF-NT technique compared with ZI-NT and ICI-NT. Cleavage rate was not affected by the SCNT protocol, whereas blastocyst yield rate in ICI-NT technique (17.0±1.0%) was significantly (P<0.05; ANOVA) higher than in ZF-NT (7.1±1.5%) but not in the ZI-NT group (11.2±3.3%). Despite the similarities in total cell number, SCNT protocol changed the distribution of cells in the blastocysts, as ZF-NT-cloned blastocysts had significantly smaller inner cell mass than ZI-NT. These results indicate that technical aspects of cloning may result in the variety of cloning phenotypes. © 2013 Society for Reproduction and Fertility.

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