Publications by authors named "Maryam Pashaiasl"

21 Publications

  • Page 1 of 1

Design and Microfabrication of An On-Chip Oocyte Maturation System for Reduction of Apoptosis.

Cell J 2021 Apr 1;23(1):32-39. Epub 2021 Mar 1.

Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Email:

Objective: In customary assisted reproductive technology (ART), oocyte culture occurs in static micro drops of Petri dishes with vast media volume; while, the condition is dynamic. In this study, we aimed to improve the maturation efficiency of mammalian oocytes by designing an optimal microchamber array to obtain the integration of oocyte trapping and maturation within a microfluidic device and evaluate the role of microfluidic culture condition in lipid peroxidation level of the culture medium, matured oocytes apoptosis, and its comparison with the conventional static system.

Materials And Methods: In this experimental research, immature oocytes were collected from ovaries of the Naval Medical Research Institute (NMRI) mice. Oocytes were randomly laid in static and dynamic (passive and active) maturation culture medium for 24 hours. The lipid peroxidation level in oocyte culture media was assessed by measuring the concentration of malondialdehyde (MDA), and the rate of apoptosis in matured oocytes was assessed by the TUNEL assay after a-24 hour maturation period.

Results: The MDA concentration in both dynamic oocyte maturation media were significantly lower than the static medium (0.003 and 0.002 vs. 0.13 μmol/L, P<0.01). Moreover, the rate of apoptosis in matured oocytes after a-24 hour maturation period was significantly lower in passive dynamic and active dynamic groups compared with the static group (16%, 15% vs. 35%, P<0.01).

Conclusion: The dynamic culture for oocyte maturation (IVM) improves the viability of IVM oocytes in comparison with the static culture condition.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.22074/cellj.2021.7056DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7944125PMC
April 2021

Investigation of molecular cryopreservation, fertility potential and microRNA-mediated apoptosis in Oligoasthenoteratozoospermia men.

Cell Tissue Bank 2021 Mar 15;22(1):123-135. Epub 2020 Oct 15.

Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Investigation of the cryo-injury mechanism can provide novel insight into cryopreservation. The objective of this study is to assess the effect of cryopreservation on fertility potential, motility, oxidative stress (OS), DNA fragmentation, microRNAs (miRNAs), and apoptotic target genes in the infertile men compared to the fertile men. All 40 samples were divided into two leading groups of fresh and cryopreserved sperms. Each main group was subdivided into three groups including, Normozoospermia, and Mild, and Severe Oligoasthenoteratozoospermia (OAT). In all collected samples the following were assessed: microRNA-34c (miR-34c) and miR-184, P53 and Caspase9 using Quantitative real-time polymerase chain reaction (RT-PCR), malondialdehyde (MDA), Superoxide dismutase (SOD) using imaging multi-mode reader, and DNA fragmentation using Sperm DNA Fragmentation Assay Test (SDFA). Within the studied groups, immotile spermatozoa were increased due to cryopreservation. We observed an increasing levels of SOD, MDA, and DNA fragmentation. Also, cryopreservation was associated with decreasing the expression of P53, mir-43c, and miR-184 while capase9 was showed enhancing expression after freeze-thawing of sperm cells. During cryopreservation, sperm fertility and motility were influenced via apoptosis cascade-mediated mitochondrial dysfunctions such as caspase9. Also, we found that miR-34c, miR184, and P53 could impact fertility potential. In Addition, there was a meaningful correlations between microRNAs and motility post freeze-thawing process in Severe Oligoasthenoteratozoospermia men.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10561-020-09872-xDOI Listing
March 2021

Bidirectional and Opposite Effects of Naïve Mesenchymal Stem Cells on Tumor Growth and Progression.

Adv Pharm Bull 2019 Oct 24;9(4):539-558. Epub 2019 Oct 24.

Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Cancer has long been considered as a heterogeneous population of uncontrolled proliferation of different transformed cell types. The recent findings concerning tumorigeneses have highlighted the fact that tumors can progress through tight relationships among tumor cells, cellular, and non-cellular components which are present within tumor tissues. In recent years, studies have shown that mesenchymal stem cells (MSCs) are essential components of non-tumor cells within the tumor tissues that can strongly affect tumor development. Several forms of MSCs have been identified within tumor stroma. Naïve (innate) mesenchymal stem cells (N-MSCs) derived from different sources are mostly recruited into the tumor stroma. N-MSCs exert dual and divergent effects on tumor growth through different conditions and factors such as toll-like receptor priming (TLR-priming), which is the primary underlying causes of opposite effects. Moreover, MSCs also have the contrary effects by various molecular mechanisms relying on direct cellto- cell connections and indirect communications through the autocrine, paracrine routes, and tumor microenvironment (TME). Overall, cell-based therapies will hold great promise to provide novel anticancer treatments. However, the application of intact MSCs in cancer treatment can theoretically cause adverse clinical outcomes. It is essential that to extensively analysis the effective factors and conditions in which underlying mechanisms are adopted by MSCs when encounter with cancer. The aim is to review the cellular and molecular mechanisms underlying the dual effects of MSCs followed by the importance of polarization of MSCs through priming of TLRs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.15171/apb.2019.063DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6912184PMC
October 2019

Influence of cryopreservation on structure and function of mammalian spermatozoa: an overview.

Cell Tissue Bank 2020 Mar 3;21(1):1-15. Epub 2019 Dec 3.

Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Cryopreservation is a useful approach to preserve male fertility for assisted reproduction technique and other evaluations. In spite of extensive development in the cryopreservation field, there are biological and biochemical points including DNA fragmentation and antioxidant which should be illuminated to preserve fertility in safe form. Several molecular markers such as coding and noncoding RNAs are transferred from spermatozoa into oocyte via fertilization. These biomarkers affect fertility potential during the cryopreservation. Despite its impact, sperm cryopreservation has some destructive effect including loss of sperm motility and viability, acrosomal damage, mitochondrial membrane depolarization, plasma membrane permeability alteration even nuclear, and DNA damage. There is a growing concern about the impact of sperm cryopreservation on biological factors which can interrupt successful fertility procedures such as in vitro fertilization. Additionally, cryo-damage can decrease embryonic development extent. Promoting cryopreservation method via investigating the wide range of non-invasive factors may be increasingly important to have access to safe approach of freezing. Therefore, the aim of this study is the assessment of biological factors which can penetrate the fertility potential during the freezing procedure, explicate innovative methods in fertility preservation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s10561-019-09797-0DOI Listing
March 2020

MicroRNA-based regulatory circuit involved in sperm infertility.

Andrologia 2020 Feb 24;52(1):e13453. Epub 2019 Nov 24.

Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

miRNAs (MicroRNAs), known as noncoding and important endogenous factors regulating the expression protein-coding genes, are vital regulators in each biological process. Thus, this study aims to explore the key role of four microRNAs in regulating the spermatogenesis. To conduct this experiment, 55 infertile and fertile men provided the study with the sperm and testicular tissue samples. To study the spermatozoa in terms of the morphology, Diff-Quick was applied. Then, quantitative real-time polymerase chain reaction (RT-PCR) was conducted on samples. Our data indicated that in contrast to the miR-15b, significant increasing of miR-383 and miR-122 occurred in both severe oligoasthenoteratozoospermia (SOAT) and moderate oligoasthenoteratozoospermia (MOAT) compared to normal sperm group (N). In addition, it was observed that miR-15b and miR-122 increased in patients with nonobstructive azoospermia (NOA) compared with obstructive azoospermia (OA) group. Expression levels of target genes including P53, CASPASE-9 and CYCLIN D1 underwent principle changes according to miRNAs expression level. Our finding indicated that miRNAs had essential role in the regulation of spermatogenesis, and their expression altering was associated with sperm abnormalities. Thus, microRNAs can be introduced as useful biomarkers to determine male infertility reasons to choose the effective treatment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/and.13453DOI Listing
February 2020

Application of nanomaterials in three-dimensional stem cell culture.

J Cell Biochem 2019 11 30;120(11):18550-18558. Epub 2019 Jul 30.

Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Petri dish cultured cells have for long provided scientists an aperture to understanding cell's behavior both in normal and disease states as well as in vitro and in vivo. But recent advances have brought to light how the architecture and composite nature of the immediate environment within which the cell is proliferated can profoundly influence its phenotypic features and functions, thus making obvious, limitations of the conventional two-dimensional cell culture despite it cost effectiveness. Fortunately, the transition to three-dimensional (3D) cell culture has occurred concurrently with expanded knowledge of nanoscience and materials, thereby lending significant impetus for innovative research. This review is focused on the application of nanoparticles in 3D stem cell breeding, recent trends and developments in medical sciences for improved drug delivery, and treatment approaches to some human diseases. We also reviewed prevailing challenges and concerns of nanotoxicity as it continues to impede and delay clinical applications as well the ongoing concerted and multidisciplinary efforts to overcome them.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jcb.29133DOI Listing
November 2019

The human amniotic fluid mesenchymal stem cells therapy on, SKOV3, ovarian cancer cell line.

Mol Genet Genomic Med 2019 07 20;7(7):e00726. Epub 2019 May 20.

Women's Reproductive Health Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Purpose: One of the most common malignancies peculiar to female health with few symptoms, low response to therapy, difficult diagnosis, frequent relapse, and high mortality, is ovarian cancer. Thus, our experiment, using Human amniotic fluid mesenchymal stem cells (hAFMSCs) as a therapeutic tool, aims to find an efficient treatment approach for patients suffering from SKOV3 ovarian cancer.

Material & Methods: In this study, we obtained 5 ml amniotic fluid from 16-20 week pregnant women who underwent amniocentesis for routine prenatal diagnosis by karyotyping in Al-Zahra Hospital of Tabriz University of Medical Sciences, Iran. Using trans wells in 24 wells plate, hAFMSCs were isolated from all samples, co-cultured with SKOV3 ovarian cancer cell line, and characterized via flow cytometry and RT-PCR. Human skin fibroblast cells (HSFCs) were isolated and used as a negative control. SKOV3 and HSFCs' viability after 5 days was evaluated by MTT assay. Cell cycle and apoptotic genes were analyzed by real-time PCR.

Results: We successfully isolated and characterized hAFMSCs through it positivity for CD44 and CD90 specific mesenchymal stem cell markers and negativity for CD31 and CD45. Oct4 and NANOG were evaluated by RT-PCR as pluripotency markers, and visualized on 2% gel electrophoresis. We established hAFMS cell lines after 5 days of co-culturing the SKOV3 cells, viability was decreased; however, HSFCs did not show toxicity by MTT assay. The genes indicated upregulation and high expression by a real-time PCR.

Conclusions: Our findings showed that hAFMSCs have natural tumor tropism, and can release soluble factors in a cell culture, which cause an efficient anticancer effect. Thus, we can use hAFMSCs for complete anticancer therapy on SKOV3 cell line at cell culture condition and possibly in vivo in the near future.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/mgg3.726DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6625370PMC
July 2019

Human amniotic fluid stem cells (hAFSCs) expressing p21 and cyclin D1 genes retain excellent viability after freezing with (dimethyl sulfoxide) DMSO.

Bosn J Basic Med Sci 2019 Feb 12;19(1):43-51. Epub 2019 Feb 12.

Department of Biochemistry, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.

Human amniotic fluid stem cells (hAFSCs) have features intermediate between embryonic and adult SCs, can differentiate into lineages of all three germ layers, and do not develop into tumors in vivo. Moreover, hAFSCs can be easily obtained in routine procedures and there is no ethical or legal limitations regarding their use for clinical and experimental applications. The aim of this study was to assess the effect of slow freezing/thawing and two different concentrations of DMSO (10% DMSO + 90% fetal bovine serum [FBS] and 5% DMSO + 95% FBS) on the survival of hAFSCs. hAFSCs were obtained from 5 pregnant women during amniocentesis at 16-22 weeks of gestation. The expression of pluripotency markers (Octamer-binding transcription factor 4 [Oct4] and NANOG) by reverse transcription polymerase chain reaction and cell surface markers (cluster of differentiation [CD31], CD44, CD45, and CD90) by flow cytometry was analyzed before and after the slow-freezing. Cell viability was assessed by trypan blue exclusion or MTT assay. Quantitative mRNA expression of Oct4, NANOG, cyclin D1 and p21 was determined by real-time PCR before and after the slow-freezing. Pluripotency of hAFSCs was confirmed by NANOG and POU5F1 (Oct4) gene expression before and after slow-freezing. All hAFSC cultures were positive for CD44 and CD90. A higher viability of hAFSCs was observed after freezing with 90% FBS + 10% DMSO. There was increased expression of NANOG and decreased expression of POU5F1 gene after freezing, compared to control cells (before freezing). DMSO and the process of freezing did not significantly change the expression of p21 and cyclin D1 genes in hAFSCs. Overall, our results indicate the applicability of slow-freezing and DMSO in cryopreservation of SCs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.17305/bjbms.2018.2912DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6387673PMC
February 2019

Enhancement of anticancer activity by silibinin and paclitaxel combination on the ovarian cancer.

Artif Cells Nanomed Biotechnol 2018 Nov 8;46(7):1483-1487. Epub 2017 Sep 8.

e Drug Applied Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.

Background: Ovarian carcinoma is the most lethal cancer among all gynaecological malignancies. One of the most chemotherapy drugs used for ovarian cancer is paclitaxel which induces apoptosis. Paclitaxel has been used for many years. Similar to the most cancers this responds to chemotherapy initially but in a long run, drug resistance happens which fails the treatment procedure. Combination of chemotherapy drugs has been suggested to deal with this issue. Silibinin, a plant extraction, has been used from ancient time in traditional medicine and identified to have powerful antioxidant activity.

Aim: The aim of this study was to examine the effect of paclitaxel and silibinin combination on SKOV-3 cancer cell line.

Materials And Methods: The human epithelial ovarian cancer cell line, SKOV-3, was cultured and treated with paclitaxel, silibinin and paclitaxel plus silibinin for 48 hours. MTT assay was carried out to determine cell viability. For apoptotic process, we used real-time PCR to study P53 and P21 genes expression after drug treatment and network analysis was performed using Pathway Studio web tool (Elsevier).

Results: Cell growth was inhibited considerably (p < .05) by combination of paclitaxel and silibinin after 48 hours of treatment. Also silibinin and paclitaxel combination induced apoptosis in SKOV-3 cells. Expression analysis by real-time PCR showed the significant up-regulation of two tumour suppressor genes, P53 and P21 in response to combination of silibinin and paclitaxel. In addition, computational network analysis demonstrated the crosstalk between paclitaxel, silibinin and ovarian cancer.

Conclusions: Our results showed that combination of chemotherapy drugs of silibinin and paclitaxel can be more efficient in treatment of ovarian cancer cells.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/21691401.2017.1374281DOI Listing
November 2018

The Inhibitory Effect of Ginger Extract on Ovarian Cancer Cell Line; Application of Systems Biology.

Adv Pharm Bull 2017 Jun 30;7(2):241-249. Epub 2017 Jun 30.

Women's Reproductive Health Research Center, Tabriz University of Medical Sciences Tabriz, Iran.

Ginger is a natural compound with anti-cancer properties. The effects of ginger and its mechanism on ovarian cancer and its cell line model, SKOV-3, are unclear. In this study, we have evaluated the effect of ginger extract on SKOV-3. SKOV-3 cells were incubated with ginger extract for 24, 48 and 72 hours. Cell toxicity assay was performed. Different data mining algorithms were applied to highlight the most important features contributing to ginger inhibition on the SKOV-3 cell proliferation. Moreover, Real-Time PCR was performed to assay p53, p21 and bcl-2 genes expression. For co-expression meta-analysis of p53, mutual ranking (MR) index and transformation to Z-values (Z distribution) were applied on available transcriptome data in NCBI GEO data repository. The ginger extract significantly inhibited cancer growth in ovarian cancer cell line. The most important attribute was 60 µg/ml concentration which received weights higher than 0.50, 0.75 and 0.95 by 90%, 80% and 50% of feature selection models, respectively. The expression level of p53 was increased sharply in response to ginger treatment. Systems biology analysis and meta-analysis of deposited expression value in NCBI based on rank of correlation and Z-transformation approach unraveled the key co-expressed genes and co-expressed network of P53, as the key transcription factor induced by ginger extract. High co-expression between P53 and the other apoptosis-inducing proteins such as CASP2 and DEDD was noticeable, suggesting the molecular mechanism underpinning of ginger action. We found that the ginger extract has anticancer properties through p53 pathway to induce apoptosis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.15171/apb.2017.029DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5527238PMC
June 2017

and Dendrimers-Mediated Antisense Therapy.

Adv Pharm Bull 2017 Jun 30;7(2):179-187. Epub 2017 Jun 30.

Department of Biology, Payame Noor University, Tehran, Iran.

Finding novel and effective antibiotics for treatment of is a challenging field. Treatment with antibiotics usually cures infection; however, if the resultant disease is not timely recognized and treated properly, it leads to poor prognosis and high case fatality rate. DrrA protein (Defects in Rab1 recruitment protein A)/also known as SidM affects host cell vesicular trafficking through modification of the activity of cellular small guanosine triphosphatase )GTPase( Rab (Ras-related in brain) function which facilitates intracellular bacterial replication within a supporter vacuole. Also, LepA and LepB (Legionella effector protein A and B) proteins suppress host-cell Rab1 protein's function resulting in the cell lysis and release of bacteria that subsequently infect neighbour cells. Legionella readily develops resistant to antibiotics and, therefore, new drugs with different modes of action and therapeutic strategic approaches are urgently required among antimicrobial drug therapies;gene therapy is a novel approach for treatment. On the contrary to the conventional treatment approaches that target bacterial proteins, new treatment interventions target DNA (Deoxyribonucleic acid), RNA (Ribonucleic acid) species, and different protein families or macromolecular complexes of these components. The above approaches can overcome the problems in therapy of Legionella infections caused by antibiotics resistance pathogens. Targeting Legionella genes involved in manipulating cellular vesicular trafficking using a dendrimer-mediated antisense therapy is a promising approach to inhibit bacterial replication within the target cells.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.15171/apb.2017.022DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5527231PMC
June 2017

Fluorescent multi-responsive cross-linked P(N-isopropylacrylamide)-based nanocomposites for cisplatin delivery.

Drug Dev Ind Pharm 2017 Aug 18;43(8):1283-1291. Epub 2017 Apr 18.

e Drug Applied Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.

Magnetic, pH and temperature-sensitive, poly(N-isopropylacrylamide) (PNIPAM)-based nanocomposites with fluorescent properties were synthesized by free radical copolymerization-cross linking of NIPAM, N,N-dimethylaminoethyl methacrylate (DMAEMA) and 4-acrylamidofluorescein (AFA). The model anti-cancer drug, cisplatin (CDDP), was loaded into the resulted nanogel. For the production of CDDP-loaded nanocomposite, FeO magnetic nanoparticles (MNPs) and CDDP were loaded into the nanogel. Field-emission scanning electron microscopy (FE-SEM) indicated that the size of nanogel and CDDP-loaded nanocomposite were about 90 and 160 nm, respectively. The encapsulation efficiency of CCDP was found up to 65%. The loaded CCDP showed sustained thermal and pH-responsive drug release. A high level of drug release was observed under the conditions of low pH and high temperature. The lower critical solution temperature (LCST) of synthesized nanogel was about 40 °C. CDDP-loaded nanocomposite showed a volume phase transition from 282 to 128 nm at its LCST. Accordingly, in this study, the synthesized nanocomposite can be employed as a stimuli-responsive anti-cancer drug delivery system and the pH and temperature of solution have the potential to monitor the drug release.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/03639045.2017.1313859DOI Listing
August 2017

An update clinical application of amniotic fluid-derived stem cells (AFSCs) in cancer cell therapy and tissue engineering.

Artif Cells Nanomed Biotechnol 2017 Jun 10;45(4):765-774. Epub 2016 Aug 10.

a Women?s Reproductive Health Research Center , Tabriz University of Medical Sciences , Tabriz , Iran.

Recent studies have elucidated that cell-based therapies are promising for cancer treatments. The human amniotic fluid stem (AFS) cells are advantageous cells for such therapeutic schemes that can be innately changed to express therapeutic proteins. HAFSCs display a natural tropism to cancer cells in vivo. They can be useful in cancer cells targeting. Moreover, they are easily available from surplus diagnostic samples during pregnancy and less ethical and legal concern are associated with the collection and application than other putative cells are subjected. This review will designate representatives of amniotic fluid and stem cell derived from amniotic fluid. For this propose, we collect state of human AFS cells data applicable in cancer therapy by dividing this approach into two main classes (nonengineered and engineered based approaches). Our study shows the advantage of AFS cells over other putative cells types in terms differentiation ability to a wide range of cells by potential and effective use in preclinical studies for a variety of diseases. This study has shown the elasticity of human AFS cells and their favorable potential as a multipotent cell source for regenerative stem cell therapy and capable of giving rise to multiple lineages including such as osteoblasts and adipocyte.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/21691401.2016.1216857DOI Listing
June 2017

Evaluation of Mouse Oocyte In Vitro Maturation Developmental Competency in Dynamic Culture Systems by Design and Construction of A Lab on A Chip Device and Its Comparison with Conventional Culture System.

Cell J 2016 Jul-Sep;18(2):205-13. Epub 2016 May 30.

Cellular and Molecular Biology Research Centre, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Objective: In conventional assisted reproductive technology (ART), oocytes are cultured in static microdrops within Petri dishes that contain vast amounts of media. However, the in vivo environment is dynamic. This study assesses in vitro oocyte maturation through the use of a new microfluidic device. We evaluate oocyte fertilization to the blastocyct stage and their glutathione (GSH) contents in each experimental group.

Materials And Methods: In this experimental study, we established a dynamic culture condition. Immature oocytes were harvested from ovaries of Naval Medical Research Institute (NMRI) mice. Oocytes were randomly placed in static (passive) and dynamic (active) in vitro maturation (IVM) culture medium for 24 hours. In vitro matured oocytes underwent fertilization, after which we placed the pronucleus (PN) stage embryos in microdrops and followed their developmental stages to blastocyst formation after 3 days. GSH content of the in vitro matured oocytes was assessed by monochlorobimane (MCB) staining.

Results: We observed significantly higher percentages of mature metaphase II oocytes (MII) in the passive and active dynamic culture systems (DCS) compared to the static group (P<0.01). There were significantly less mean numbers of germinal vesicle (GV) and degenerated oocytes in the passive and active dynamic groups compared to the static group (P<0.01). Fertilization and blastocyst formation rate in the dynamic systems were statistically significant compared to the static cultures (P<0.01). There was significantly higher GSH content in dynamically matured oocytes compared to statically matured oocytes (P<0.01).

Conclusion: Dynamic culture for in vitro oocyte maturation improves their developmental competency in comparison with static culture conditions.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4988419PMC
http://dx.doi.org/10.22074/cellj.2016.4315DOI Listing
August 2016

Isolation, Characterization, Cryopreservation of Human Amniotic Stem Cells and Differentiation to Osteogenic and Adipogenic Cells.

PLoS One 2016 19;11(7):e0158281. Epub 2016 Jul 19.

Women's Reproductive Health Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Human stem cells and progenitor cells can be used to treat cancer and replace dysfunctional cells within a tissue or organ. The objective of this study was to identify the appropriate cells type in regenerative medicine and targeted therapy. As an alternative to embryonic and bone marrow stem cells, we examined human amniotic fluid stem cells (hAFSCs), one of the potential source of multipotent stem cells isolated from both cell pellet (using single-stage method), and supernatant of human amniotic fluid. Source of isolation and unique property of the cells emphasize that these cells are one of the promising new tools in therapeutic field. Double sources for isolation and availability of the left over samples in diagnostic laboratory at the same time have less legal and ethical concerns compared with embryonic stem cell studies. Cells were isolated, cultured for 18th passage for 6 months and characterized using qPCR and flow cytometry. Cells showed good proliferative ability in culture condition. The cells successfully differentiated into the adipogenic and osteogenic lineages. Based on these findings, amniotic fluid can be considered as an appropriate and convenient source of human amniotic fluid stem cells. These cells provide potential tools for therapeutic applications in the field of regenerative medicine. To get a better understanding of crosstalk between Oct4/NANOG with osteogenesis and adipogenesis, we used network analysis based on Common Targets algorithm and Common Regulators algorithm as well as subnetwork discovery based on gene set enrichment. Network analysis highlighted the possible role of MIR 302A and MIR let-7g. We demonstrated the high expression of MIR 302A and low expression of MIR let7g in hAFSCs by qPCR.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0158281PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4951121PMC
July 2017

Identification of the key regulating genes of diminished ovarian reserve (DOR) by network and gene ontology analysis.

Mol Biol Rep 2016 Sep 20;43(9):923-37. Epub 2016 Jun 20.

Division of Information Technology, Engineering and the Environment, School of Information Technology and Mathematical Sciences, University of South Australia, Adelaide, Australia.

Diminished ovarian reserve (DOR) is one of the reasons for infertility that not only affects both older and young women. Ovarian reserve assessment can be used as a new prognostic tool for infertility treatment decision making. Here, up- and down-regulated gene expression profiles of granulosa cells were analysed to generate a putative interaction map of the involved genes. In addition, gene ontology (GO) analysis was used to get insight intol the biological processes and molecular functions of involved proteins in DOR. Eleven up-regulated genes and nine down-regulated genes were identified and assessed by constructing interaction networks based on their biological processes. PTGS2, CTGF, LHCGR, CITED, SOCS2, STAR and FSTL3 were the key nodes in the up-regulated networks, while the IGF2, AMH, GREM, and FOXC1 proteins were key in the down-regulated networks. MIRN101-1, MIRN153-1 and MIRN194-1 inhibited the expression of SOCS2, while CSH1 and BMP2 positively regulated IGF1 and IGF2. Ossification, ovarian follicle development, vasculogenesis, sequence-specific DNA binding transcription factor activity, and golgi apparatus are the major differential groups between up-regulated and down-regulated genes in DOR. Meta-analysis of publicly available transcriptomic data highlighted the high coexpression of CTGF, connective tissue growth factor, with the other key regulators of DOR. CTGF is involved in organ senescence and focal adhesion pathway according to GO analysis. These findings provide a comprehensive system biology based insight into the aetiology of DOR through network and gene ontology analyses.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s11033-016-4025-8DOI Listing
September 2016

Unravelling evolution of Nanog, the key transcription factor involved in self-renewal of undifferentiated embryonic stem cells, by pattern recognition in nucleotide and tandem repeats characteristics.

Gene 2016 Mar 12;578(2):194-204. Epub 2015 Dec 12.

Department of Biology, University of Qom, Qom, Iran. Electronic address:

Nanog, an important transcription factor in embryonic stem cells (ESC), is the key factor in maintaining pluripotency to establish ESC identity and has the ability to induce embryonic germ layers. Nanog is responsible for self-renewal and pluripotency of stem cells as well as cancer invasiveness, tumor cell proliferation, motility and drug-resistance. Understanding the underlying mechanisms of Nanog evolution and regulation can lead to future advances in treatment of cancers. Recent integration of machine learning models with genetics has provided a powerful tool for knowledge discovery and uncovering evolutionary pathways. Herein, sequences of 47 Nanog genes from various species were extracted and two datasets of features were computationally extracted from these sequences. At the first dataset, 76 nucleotide acid attributes were calculated for each Nanog sequence. The second dataset was prepared based on the 10,480 repeated nucleotide sequences (from 5 to 50bp lengths). Then, various data mining algorithms such as decision tree models were applied on these datasets to find the evolutionary pathways of Nanog diversion. Attribute weighting models were highlighted features such as the frequencies of AA and GC as the most important genomic features in Nanog gene classification and differentiation. Similar findings were obtained by tree induction algorithms. Results from the second database showed that some short sequence strings, such as ACTACT, TCCTGA, CCTGA, GAAGAC, and TATCCC can be effectively used to identify Nanog genes in various species. The outcomes of this study, for the first time, unravels the importance of particular genomic features in Nanog gene evolution paving roads toward better understanding of stem cell development and human targeted disorder therapy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.gene.2015.12.023DOI Listing
March 2016

A Mini Overview of Isolation, Characterization and Application of Amniotic Fluid Stem Cells.

Int J Stem Cells 2015 Nov;8(2):115-20

Department of Reproductive Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Amniotic fluid represents rich sources of stem cells that can be used in treatments for a wide range of diseases. Amniotic fluid- stem cells have properties intermediate between embryonic and adult mesenchymal stem cells which make them particularly attractive for cellular regeneration and tissue engineering. Furthermore, scientists are interested in these cells because they come from the amniotic fluid that is routinely discarded after birth. In this review we give a brief introduction of amniotic fluid followed by a description of the cells present within this fluid and aim to summarize the all existing isolation methods, culturing, characterization and application of these cells. Finally, we elaborate on the differentiation and potential for these cells to promote regeneration of various tissue defects, including fetal tissue, the nervous system, heart, lungs, kidneys, bones, and cartilage in the form of table.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.15283/ijsc.2015.8.2.115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4651275PMC
November 2015

Cryopreservation and long-term maintenance of bovine embryo-derived cell lines.

Reprod Fertil Dev 2013 ;25(4):707-18

Centre for Reproduction and Development, Monash Institute of Medical Research, Clayton, Vic. 3168, Australia.

The aim of this study was to develop methods for cryopreservation and long-term maintenance of putative bovine embryonic stem cells (ESCs). Putative bovine ESC (bESC) lines (n=3) isolated in conventional medium were used to compare slow-freezing and vitrification. After warming, vitrified cells (96.9%) demonstrated significantly (P<0.05) better survival than frozen-thawed cells (81.5%) and formed significantly more colonies with good morphology (vitrification: 93/93, 100.0%; slow-freezing: 74/106, 69.81%; P<0.05). The effect of inhibitors of differentiation (PD184352, SU5402, CHIR99021) on ESC maintenance was assessed on putative bESC lines established in N2B27-3i medium (n=8) or conventional medium (n=1) after culture over 30 passages (>240 days). All cell lines expressed ALP, SSEA1, SSEA4, OCT4, REX1 and SSEA1. OCT4 expression was confirmed by relative real-time PCR and was upregulated in early passages of putative bESCs cultured in N2B27-3i (2.9±0.89-fold higher at Passage (P) 2-4), whereas the converse was observed later (P22-26; 2.2±0.1-fold increase in conventional medium). Putative bESC lines isolated in N2B27-3i medium (n=3) or conventional medium (n=1) were vitrified at P18 and, after warming, were cultured for a further 12 passages. These cells survived vitrification and expressed OCT4, REX1, SSEA1, ALP, SSEA1 and SSEA4. These results demonstrate that putative bESC lines that express pluripotent markers can be cultured long term and retain expression of pluripotent markers after vitrification.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1071/RD12018DOI Listing
November 2013

Induction of pluripotency in adult equine fibroblasts without c-MYC.

Stem Cells Int 2012 19;2012:429160. Epub 2012 Mar 19.

Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, Clayton, VIC 3800, Australia.

Despite tremendous efforts on isolation of pluripotent equine embryonic stem (ES) cells, to date there are few reports about successful isolation of ESCs and no report of in vivo differentiation of this important companion species. We report the induction of pluripotency in adult equine fibroblasts via retroviral transduction with three transcription factors using OCT4, SOX2, and KLF4 in the absence of c-MYC. The cell lines were maintained beyond 27 passages (more than 11 months) and characterized. The equine iPS (EiPS) cells stained positive for alkaline phosphatase by histochemical staining and expressed OCT4, NANOG, SSEA1, and SSEA4. Gene expression analysis of the cells showed the expression of OCT4, SOX2 NANOG, and STAT3. The cell lines retained a euploid chromosome count of 64 after long-term culture cryopreservation. The EiPS demonstrated differentiation capacity for the three embryonic germ layers both in vitro by embryoid bodies (EBs) formation and in vivo by teratoma formation. In conclusion, we report the derivation of iPS cells from equine adult fibroblasts and long-term maintenance using either of the three reprogramming factors.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1155/2012/429160DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3328202PMC
July 2012

The efficient generation of cell lines from bovine parthenotes.

Cell Reprogram 2010 Oct;12(5):571-9

Centre for Reproduction and Development, Monash Institute of Medical Research, VIC, Australia.

The generation of embryonic stem cell (ESC) lines from parthenogenetically activated oocytes can provide transplantable cells, which are immunocompatible for the oocyte donors as well as an invaluable tool for genetic engineering and epigenetic studies. We report the efficient isolation of eight putative bovine parthenogenetic embryonic stem cell (bpESC) lines from 15 in vitro produced parthenotes. Five of these cell lines were maintained for more than 15 passages (>140 days) and analyzed. The cells displayed typical ESC morphology, stained positive for alkaline phosphate by histochemical staining, expressed Oct4, Nanog, and either stage-specific embryonic antigens, SSEA1, or SSEA4, detected by immunofluorescence staining. RT-PCR analysis of the cells demonstrated expression of Oct4, Rex1, SSEA1, and ALP. All the cell lines except one had a normal karyotype of 60, XX. The cells differentiated in suspension culture to form embryoid bodies (EBs) expressing markers of the three embryonic germ layers as assessed by RT-PCR. In conclusion, we report efficient derivation of putative ESCs from bovine parthenogenetic embryos. The cells express pluripotent markers, have the ability to form EBs, and differentiate into cells of the three embryonic germ layers. This is the first report of characterized putative parthenogenetic bovine ESC lines.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1089/cell.2009.0118DOI Listing
October 2010