ABSTRACT
Objective: Three-dimensional [3D] biomimetic nanofiber scaffolds have widespread applications in biomedical tissue engineering. They provide a suitable environment for cellular adhesion, survival, proliferation and differentiation, guide new tissue formation and development, and are one of the outstanding goals of tissue engineering. Electrospinning has recently emerged as a leading technique for producing biomimetic scaffolds with micro to nanoscale topography and a high porosity similar to the natural extracellular matrix [ECM]. These scaffolds are comprised of synthetic and natural polymers for tissue engineering applications. Several kinds of cells such as human embryonic stem cells [hESCs] and mouse ESCs [mESCs] have been cultured and differentiated on nanofiber scaffolds. mESCs can be induced to differentiate into a particular cell lineage when cultured as embryoid bodies [EBs] on nano-sized scaffolds
Materials and Methods: We cultured mESCs [2500 cells/100 Mul] in 96-well plates with knockout Dulbecco's modified eagle medium [DMEM-KO] and Roswell Park Memorial Institute-1640 [RPMI-1640], both supplemented with 20% ESC grade fetal bovine serum [FBS] and essential factors in the presence of leukemia inhibitory factor [LIF]. mESCs were seeded at a density of 2500 cells/100 Mul onto electrospun polycaprolactone [PCL] nanofibers in 96-well plates. The control group comprised mESCs grown on tissue culture plates [TCP] at a density of 2500 cells/100 Mul. Differentiation of mESCs into mouse hematopoietic stem cells [mHSCs] was performed by stem cell factor [SCF], interleukin-3 [IL-3], IL-6 and Fms-related tyrosine kinase ligand [Flt3-L] cytokines for both the PCL and TCP groups. We performed an experimental study of mESCs differentiation
Results: PCL was compared to conventional TCP for survival and differentiation of mESCs to mHSCs. There were significantly more mESCs in the PCL group. Flowcytometric analysis revealed differences in hematopoietic differentiation between the PCL and TCP culture systems. There were more CD34+ [Sca1+] and CD133+ cells subpopulations in the PCL group compared to the conventional TCP culture system
Conclusion: The nanofiber scaffold, as an effective surface, improves survival and differentiation of mESCs into mHSCs compared to gelatin coated TCP. More studies are necessary to understand how the topographical features of electrospun fibers affect cell growth and behavior. This can be achieved by designing biomimetic scaffolds for tissue engineering
Materials and Methods: We cultured mESCs [2500 cells/100 Mul] in 96-well plates with knockout Dulbecco's modified eagle medium [DMEM-KO] and Roswell Park Memorial Institute-1640 [RPMI-1640], both supplemented with 20% ESC grade fetal bovine serum [FBS] and essential factors in the presence of leukemia inhibitory factor [LIF]. mESCs were seeded at a density of 2500 cells/100 Mul onto electrospun polycaprolactone [PCL] nanofibers in 96-well plates. The control group comprised mESCs grown on tissue culture plates [TCP] at a density of 2500 cells/100 Mul. Differentiation of mESCs into mouse hematopoietic stem cells [mHSCs] was performed by stem cell factor [SCF], interleukin-3 [IL-3], IL-6 and Fms-related tyrosine kinase ligand [Flt3-L] cytokines for both the PCL and TCP groups. We performed an experimental study of mESCs differentiation
Results: PCL was compared to conventional TCP for survival and differentiation of mESCs to mHSCs. There were significantly more mESCs in the PCL group. Flowcytometric analysis revealed differences in hematopoietic differentiation between the PCL and TCP culture systems. There were more CD34+ [Sca1+] and CD133+ cells subpopulations in the PCL group compared to the conventional TCP culture system
Conclusion: The nanofiber scaffold, as an effective surface, improves survival and differentiation of mESCs into mHSCs compared to gelatin coated TCP. More studies are necessary to understand how the topographical features of electrospun fibers affect cell growth and behavior. This can be achieved by designing biomimetic scaffolds for tissue engineering
ABSTRACT
Background: there are conflicting findings about relationship between depression and anger with immunological parameters
Objective: to investigate the relationship between anger patterns and immune system in depressed patients
Methods: thirty-five patients with major depressive disorder were selected according to DSM-IV criteria. The Hamilton Depression Scale and Spielberger Anger questionnaires were used to determine severity of depression and "anger expression pattern", respectively. The control group without a previous history of mental illness was also selected. In the group of patients with moderate depression, serum IgA levels and NK cell percentage were measured
Results: mean differences of all types of "anger expression pattern", including; "state-trait anger", "anger expression out", "anger expression in", "anger control out" and "anger control in", between study and control groups, were statistically significant [p<0.05]. Difference in mean serum levels of IgA in either group was not significant [p=0.9], but the mean difference was significant in terms of NK-cell percentage in both groups [p=0.04]. There was no significant relationship between IgA levels and percentage of NK-cell with all types of "anger expression pattern" in both groups. Only in the control group, IgA had significant correlation with Anger control out [p=0.04]
Conclusion: moderately depressed patients versus control group had higher Spielberger scores in all types of anger expression pattern except anger controlout and anger control-in. We found no evidence supporting the relationship between" anger expression pattern" and IgA levels and NK cell percentage; however, it seems that depression itself causes reduced number of NK cells and increased IgA levels
ABSTRACT
Objective: Detection of chromosomal translocations has an important role in diagnosis and treatment of hematological disorders. We aimed to evaluate the 46 new cases of de novo acute myeloid leukemia [AML] patients for common translocations and to assess the effect of geographic and ethnic differences on their frequencies
Materials and Methods: In this descriptive study, reverse transcriptase-polymerase chain reaction [RT-PCR] was used on 46 fresh bone marrow or peripheral blood samples to detect translocations t [8; 21], t [15; 17], t [9; 11] and inv [16]. Patients were classified using the French-American-British [FAB] criteria in to eight sub-groups [M0-M7]. Immunophenotyping and biochemical test results of patients were compared with RT-PCR results
Results: Our patients were relatively young with a mean age of 44 years. AML was relatively predominant in female patients [54.3%] and most of patients belonged to AML-M2. Translocation t [8; 21] had the highest frequency [13%] and t [15; 17] with 2.7% incidence was the second most frequent. CD19 as an immunophenotypic marker was at a relatively high frequency [50%] in cases with t [8; 21] and patients with this translocation had a specific immunophenotypic pattern of complete expression of CD45, CD38, CD34, CD33 and HLA-DR
Conclusion: Similarities and differences of results in Iran with different parts of the world can be explained with ethnic and geographic factors in characterizations of AML. Recognition of these factors especially in other comprehensive studies may aid better diagnosis and management of this disease
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Translocation, Genetic , Geography , Ethnicity , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Superparamagnetic iron oxide nanoparticles [SPIONs] have been used to label mammalian cells and to monitor their fate in vivo using magnetic resonance imaging [MRI]. However, the effectiveness of phenotype of labeled cells by SPIONs is still a matter of question. The aim of this study was to investigate the efficiency and biological effects of labeled mouse embryonic stem cells [mESCs] using ferumoxide- protamine sulfate complex. In an experimental study, undifferentiated mESCs, C571 line, a generous gift of Stem Cell Technology Company, were cultured on gelatin-coated flasks. The proliferation and viability of SPION-labeled cells were compared with control. ESCs and embryoid bodies [EBs] derived from differentiated hematopoietic stem cells [HSCs] were analyzed for stage-specific cell surface markers using fluorescence-activated cell sorting [FACS]. Our observations showed that SPIONs have no effect on the self-renewal ability of mESCs. Reverse microscopic observations and prussian blue staining revealed 100% of cells were labeled with iron particles. SPION-labeled mESCs did not significantly alter cell viability and proliferation activity. Furthermore, labeling did not alter expression of representative surface phenotypic markers such as stage-specific embryonic antigen 1 [SSEA1] and cluster of differentiation 117 [CD117] on undifferentiated ESC and CD34, CD38 on HSCs, as measured by flowcytometry. According to the results of the present study, SPIONs-labeling method as MRI agents in mESCs has no negative effects on growth, morphology, viability, proliferation and differentiation that can be monitored in vivo, noninvasively. Non-invasive cell tracking methods are considered as new perspectives in cell therapy for clinical use and as an easy method for evaluating the placement of stem cells after transplantation
ABSTRACT
CD34 is a type I membrane protein with a molecular mass of approximately 110 kDa. This antigen is associated with human hematopoietic progenitor cells and is a differentiation stage-specific leukocyte antigen. In this study we have generated and characterized monoclonal antibodies [mAbs] directed against a CD34 marker. Mice were immunized with two keyhole lympet hemocyanin [KLH]-conjugated CD34 peptides. Fused cells were grown in hypoxanthine, aminopterine and thymidine [HAT] selective medium and cloned by the limiting dilution [L.D] method. Several monoclones were isolated by three rounds of limited dilutions. From these, we chose stable clones that presented sustained antibody production for subsequent characterization. Antibodies were tested for their reactivity and specificity to recognize the CD34 peptides and further screened by enzyme-linked immunosorbent assay [ELISA] and Western blotting analyses. One of the mAbs [3D5] was strongly reactive against the CD34 peptide and with native CD34 from human umbilical cord blood cells [UCB] in ELISA and Western blotting analyses. The results have shown that this antibody is highly specific and functional in biomedical applications such as ELISA and Western blot assays. This monoclonal antibodies [mAb] can be a useful tool for isolation and purification of human hematopoietic stem cells [HSCs]