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1.
Cell Journal [Yakhteh]. 2019; 20 (4): 576-583
in English | IMEMR | ID: emr-199629

ABSTRACT

Objective: Hemoglobinopathies such as beta-thalassemia and sickle cell disease [SCD] are inherited disorders that are caused by mutations in beta-globin chain. Gamma-globin gene reactivation can ameliorate clinical manifestations of betathalassemia and SCD. Drugs that induce fetal hemoglobin [HbF] can be promising tools for treatment of beta-thalassemia and SCD patients. Recently, it has been shown that Simvastatin [SIM] and Romidepsin [ROM] induce HbF. SIM is a BCL11a inhibitor and ROM is a HDAC inhibitor and both of these drugs are Food and Drug Administration [FDA]-approved for hypercholesterolemia and cutaneous T-cell lymphoma respectively. Our aim was to evaluate the synergistic effects of these drugs in inducing HbF


Materials and Methods: In our experimental study, we isolated CD34+ cells from five cord blood samples that were cultured in erythroid differentiation medium containing ROM and Simvastatin. Then Gamma-globin, BCL11a and HDAC gene expression were evaluated on the 7th and 14th day of erythroid differentiation by real-time polymerase chain reaction [PCR] and immunocytochemistry


Results: Our results showed that combination of SIM and ROM significantly increased Gamma-globin gene expression and inhibit BCL11a and HDAC expression compared to results of using each of them alone. SIM and ROM lead to 3.09- fold increase in HbF production compared to the control group. Also, SIM inhibited BCL11a expression [0.065-fold] and ROM inhibited HDAC1 expression [0.47-fold] as two important inhibitors of HbF production after birth


Conclusion: We propose combination therapy of these drugs may be ameliorate clinical manifestation in beta-thalassemia and SCD with at least side effects and reduce the need for blood transfusion

2.
Cell Journal [Yakhteh]. 2019; 21 (1): 78-85
in English | IMEMR | ID: emr-203101

ABSTRACT

Objective: The role of epigenetic in regulating of the gene expression profile the embryo has been documented. MicroRNAs [miRNAs] are one of these epigenetic mechanisms. Twins are valuable models in determining the relative contributions of genetics and the environment. In this study, we compared differences in the expression levels of 44 miRNAs in hematopoietic stem cells [HSCs] of identical twins to that of fraternal twins as a controls


Materials and Methods: In this experimental study, CD133+ HSCs were isolated from cord blood of identical and fraternal twins via magnetic-activated cell sorting [MACS]. Variation in of gene expression levels of 44 miRNAs were evaluated using quantitative reverse transcription-polymerase chain reaction [qRT-PCR]


Results: Significant differences in expression were observed in both fraternal and identical twins to varying degrees, but variations alteration in expression of the miRNAs were higher in fraternal twins


Conclusion: Identical twins had a positive correlation in miRNA expression, while the correlation was not statistically significant in fraternal twins. Altogether, more differences in miRNA expression level in fraternal twins can be attributed to the both genetics and the intrauterine environment. The contribution of the intrauterine environment and genetics to miRNAs expression in HSCs was estimated 8 and 92%, respectively. By comparing of miRNA expression in identical and fraternal twins and identification of their target genes and biological pathways, it could be possible to estimate the effects of genetics and the environment on a number of biological pathways

3.
Cell Journal [Yakhteh]. 2017; 19 (Supp. 1): 27-36
in English | IMEMR | ID: emr-189337

ABSTRACT

Objective: Multiple Myeloma [MM] is a heterogeneous cytogenetic disorder in which clonal plasma cells proliferate in the bone marrow [BM] and cause bone destruction. The BM microenvironment plays a crucial role in pathogenesis of this disease, and mesenchymal stem cells [MSCs] are one of the key players. Herein, we propose to investigate the expressions of hsa-MIR-204, runt-related transcription factor 2 [RUNX2], peroxisome proliferator-activated receptor gamma [PPAR gamma], and B-cell lymphoma 2 [BCL2] as factors involved in osteogenesis, adipogenesis, and MSC survival in BM-MSCs from MM patients and normal individuals


Materials and Methods: In this experimental study, we isolated MSCs from BM aspirates of MM patients and healthy donors. Total RNA were extracted before and after co-culture with L363 myeloma cells. Gene expressions of RUNX2, PPAR gamma, BCL2, and hsa-MIR-204 were assessed by quantitive real time polymerase chain reaction [qRT-PCR]


Results: Higher levels of RUNX2, PPAR gamma, and hsa-MIR-204 expressions existed in MM-MSCs compared to normally derived [ND]-MSCs. BCL2 expression decreased in MM-MSCs. We observed different results in the co-culture model


Conclusion: In general, the MM-MSCs gene expression profile differed compared to ND-MSCs. Upregulation of RUNX2, PPAR gamma, and hsa-MIR-204 in MM-MSCs compared to ND-MSCs would result in formation of bone defects. Downregulation of BCL2 would lead to MM-MSC cell death


Subject(s)
Humans , Male , Middle Aged , Aged , Mesenchymal Stem Cells , Bone Marrow , MicroRNAs , Core Binding Factor Alpha 1 Subunit , PPAR gamma , Proto-Oncogene Proteins c-bcl-2 , Osteogenesis , Adipogenesis
4.
Journal of Tehran University Heart Center [The]. 2017; 12 (2): 65-71
in English | IMEMR | ID: emr-190813

ABSTRACT

Background: myocardial infarction is the main cause of death worldwide. Angiogenesis, a promising new therapy for the treatment of diffuse coronary artery disease, shows a poor response to conventional revascularization techniques. This study focused on improving myocardial function using endothelial cells [ECs] and mesenchymal stem cells [MSCs] in a sheep animal model


Methods: acute myocardial infarction was induced in 18 sheep [12 treated cases and 6 controls]. Autologous MSCs and ECs were injected in the infarcted area and the border zone. Two months after transplantation, echocardiography, electron microscopy, and immunohistochemistry were performed


Results: echocardiography in both MSC and EC groups revealed a significant improvement in the ejection fraction compared with the control group [p value < 0.05]. Vascular density, estimated by antibodies against the von Willebrand factor and smooth muscle actin, increased in both study groups. The pattern of vascularity in the MSC and EC groups was diffused. The electron microscopic evaluation of the infracted areas revealed cardiomyocytes in variable stages of development in the border zone in both EC and MSC groups


Conclusion: both ECs and MSCs were able to promote angiogenesis and improve cardiac function. Presumably, MSCs differentiate into ECs and cause angiogenesis as it occurs for ECs

5.
IJML-International Journal of Medical Laboratory. 2016; 3 (1): 1-12
in English | IMEMR | ID: emr-187084

ABSTRACT

Hematopoietic stem/progenitor cells [HSPCs] which give rise to different blood cell types are present within the bone marrow microenvironment, especially in flat bones such as skull, vertebrae, pelvis and chest. Interacting factors such as stromal derived factor-1/CXCR4, very late antigen-4/vascular cell adhesion molecule-1, Lymphocyte function-associated antigen-1/ intercellular adhesion molecule-1 retain the cells in the microenvironment. Any factor affecting these links may lead to migration and mobilization of HSPCs into peripheral blood. Several factors are involved in hematopoietic stem cells [HSC] mobilization such as granulocyte-colony stimulating factor, sphingosine-1-phosphate, hepatocyte growth factor, complement system, plasminogen system and matrix metalloproteinases. In bone marrow transplantation, HSC is transferred to the recipient from bone marrow of the donor, which can be performed in two ways. In the first method, Jamshidi needle is used for aspiration of bone marrow to extract hematopoietic cells usually from the hip. The second method uses mobilizer factors such as granulocyte-colony stimulating factor and granulocyte-macrophage colony-stimulating factor to mobilize the HSC into peripheral blood. Mobilized hematopoietic stem cells are suitable for the bone marrow transplantation in leukemias such as chronic myeloid leukemia, acute myeloid leukemia, chronic lymphocyte leukemia, Hairy cell leukemia, etc

6.
Archives of Medical Laboratory Sciences. 2015; 1 (3): 118-128
in English | IMEMR | ID: emr-186337

ABSTRACT

Acute lymphoblastic leukemia[ALL] is due to early stage arrest of lymphoblast development. The translocation of Philadelphia [Ph] chromosome occurs as a result of the BCR-ABL fusion gene, which constitutively produced activated tyrosine kinase. This gene fusion is an important indicator for prognosis in ALL and is associated with poor overall survival and remission duration. BCR-ABL could interfere in establishment of ALL. Therefore, in this study, we will try to investigate most pathological aspects involved in BCR-ABL fusion. Strategies for genetic alterations in B-ALL pathogenesis are discussed. Then, the main cytogenetic changes and genetic subtypes for ALL are highlighted. Moreover, intermediate reactions between cancer stem cells [CSC] related to ALL, its niche and microenvironment is discussed. The main objective in this review is to understand the principle prognosis in ALL to introduce new approaches and treatment alternatives

7.
IJML-International Journal of Medical Laboratory. 2015; 2 (3): 188-193
in English | IMEMR | ID: emr-186360

ABSTRACT

Background and Aims: iron deficiency anemia [IDA] and beta thalassemia minor [BTM] are the most common hypochromic microcytic anemias, and it is regarded important to differentiate between them. Several formulae can be taken into consideration based on red blood cell [RBC] parameters in order to distinguish between these two disorders. Hence, the present study intended to evaluate the sensitivity as well as specificity of some of these formulae


Materials and Methods: in this cross-sectional study, IDA was diagnosed in 200 patients based on hypochromic and microcytic RBC appearance, reduced mean cell volume, mean cell hemoglobin and ferritin as well as increased total iron binding capacity. Furthermore, BTM was diagnosed via hypochromic microcytic appearance of RBC and increased hemoglobin A2 level. Then, IDA and BTM diagnosis were confirmed using the formulae of King-Green, Mentzer index, England and Fraser, Shine and Lal, Srivastava and Sirdah. The sensitivity and specificity of these formulae were calculated as well


Results: the study findings demonstrated that based on the study criteria, out of 200 patients with hypochromic microcytic RBC appearance, 120 were afflicted with IDA and 80 suffered from BTM. The formulae-based diagnosis demonstrated that King- Green formula was the most reliable one


Conclusions: although King-Green formula had the highest sensitivity and specificity and was the most reliable formula, none of the formulae revealed 100% sensitivity and specificity. As a result, making definitive distinction between IDA and BTM is not possible using these formulae

8.
Cell Journal [Yakhteh]. 2014; 16 (1): 43-52
in English | IMEMR | ID: emr-148446

ABSTRACT

Finding cell sources for cartilage tissue engineering is a critical procedure. The purpose of the present experimental study was to test the in vitro efficacy of the beta-tricalcium phosphate-alginate-gelatin [BTAG] scaffold to induce chondrogenic differentiation of human umbilical cord blood-derived unrestricted somatic stem cells [USSCs]. In this experimental study, USSCs were encapsulated in BTAG scaffold and cultured for 3 weeks in chondrogenic medium as chondrogenic group and in Dulbecco's Modified Eagle's Medium [DMEM] as control group. Chondrogenic differentiation was evaluated by histology, immunofluorescence and RNA analyses for the expression of cartilage extracellular matrix components. The obtain data were analyzed using SPSS version 15. Histological and immunohistochemical staining revealed that collagen 2 was markedly expressed in the extracellular matrix of the seeded cells on scaffold in presence of chondrogenic media after 21 days. Reverse transcription-polymerase chain reaction [RT-PCR] showed a significant increase in expression levels of genes encoded the cartilage- specific markers, aggrecan, type 1 and 2 collagen, and bone morphogenetic protein [BMP]-6 in chondrogenic group. This study demonstrates that BTAG can be considered as a suitable scaffold for encapsulation and chondrogenesis of USSCs


Subject(s)
Humans , Chondrogenesis , Calcium Phosphates , Fetal Blood , Alginates , Mesenchymal Stem Cells
9.
Tehran University Medical Journal [TUMJ]. 2014; 72 (6): 367-378
in Fa | IMEMR | ID: emr-153343

ABSTRACT

Stem cells are applied in the treatment of wide range of diseases and can be separated from different tissues of the body. These cells can treat diseases by cytokine and growth factor secretion and also cell differentiation. Burn wound is a challenging problem of reconstructive surgery and stem cells may help wound healing process. We designed this study to evaluate the beneficial effect of fat derived stem cells for coverage of 3[rd] degree burn wound. This study was experimental and has been done in Burn Research Center of Iran University of Medical Sciences during January 2012 to April 2013. Thirty rats randomly divided to three equal groups. Inguinal fat of 10 rats [one group] were used for preparation of autologous adipose-derived mesenchymal stem cells. Acellular amnion was used as a scaffold for stem cell transfer. Each of the thirty rats had been exposed to a cm deep 3rd degree burn on back area. 24 hours after surgery, the wound was excised and it had been covered by three conventional dressing in the first group, acellular amnion in the second group and acellular amnion seeded with adipose-derived stem cell in the third group. The rate of wound healing and pathologic characteristics was compared in all three groups. Healing rate and decrease in wounds size was significantly better in acellular amnion seeded with adipose- derived stem cells compared with other two groups at 3[rd] and 15[th] days after surgery P<0.01. Also in histopathology examination, fibroplasia and neovascularization of wounds were significantly better in stem cells group than the other two groups P<0.001. Acellular amnion seeded with adipose-derived stem cell can result in faster wound healing and better histopathology characteristic. The amnion as a scaffold and the fat derived stem cells as healing accelerator are recommended for coverage of the 3rd degree burn wounds after excision and it may reduce the need for skin graft

10.
Modares Journal of Medical Sciences, Pathobiology. 2011; 14 (2): 1-12
in Fa | IMEMR | ID: emr-136896

ABSTRACT

The study of erythropoiesis needs to develop the methods for erythroid progenitor cells [EPCs] culture using stem cell potency to differentiate into variety of hematopoietic cells lineages. In this study, we induced differentiation of cord blood stem cells into erythroid progenitor cells in a semisolid culture media. Cord blood mononuclear cells were isolated and cultured in liquid culture media consist of erythroid differentiation factors [phase I]. Non-adherent cells were cultured in semisolid media containing SCF, IL-3, IL-6 and EPO [phase II]. After one week, the appeared erythroid colonies were picked up. Characterization of differentiated cells was performed by assessment of CD235a and CD36 using flowcytometry, giemsa cytological staining and gene expression analysis of GATA-1, EKLF, -globin genes by RT-PCR. Flowcytometry analysis to detect CD235a and CD36 positive cells showed that 94.3% and 95.5% of differentiated cells have erythroid specific cell markers, respectively. Morphology of the cells in giemsa stained slides demonstrated erythroid progenitor cells, ranged from proerythroblast to orthochromatic erythroblast. The RT-PCR results, confirmed the precursor state of erythroid differentiated cells by expression of GATA-1, EKLF, -globin genes. Cord blood stem cells have high potency to differentiate into erythroid progenitor cells using described method that can be utilized in the experimental studies

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