16 research outputs found
Selegiline Differentiates Adult Stem Cells toward Dopaminergic-Like Neurons: A Comparison between Two Cellular Niches of Hippocampal Neurogenesis
Objective: Neural stem cells (NSCs) are suitable therapeutic candidates. Here, we compare the proliferation rate,differentiation potential, and expression levels of specific markers in two groups of cultured NSCs derived from ratsubgranular (SGZ) and subventricular (SVZ) zones.Materials and Methods: In this experimental study, NSCs isolated from SGZ and SVZ were cultured in α-minimalessential medium (α-MEM) supplemented with 1% penicillin/streptomycin, 10% fetal bovine serum (FBS), 20 ng/mlbasic fibroblast growth factor (bFGF), 20 ng/ml epidermal growth factor (EGF), and B27 supplement. Glial fibrillaryacidic protein (Gfap), p75 neurotrophin receptor (Ngfr), tyrosine kinase receptor A (TrkA), beta-tubulin III (βTIII), andNestin gene levels were compared via reverse transcription polymerase chain reaction (RT-PCR) in these NSCs.Nestin and Gfap protein levels were compared by immunoassay. Subsequently, both populations were induced with10-8 M selegiline for 48 hours, followed by immunohistochemical analysis of tyrosine hydroxylase (TH) levels. One-wayANOVA and Tukey’s post-test were used with a significance level of P<0.05.Results: Both groups were successfully expanded in vitro and expressed the neurotrophin receptor genes. The SGZNSCshad a significantly higher proliferation rate and significantly higher numbers of Nestin and Gfap-positive cells.Although the majority of selegiline-induced NSCs were TH-positive, we observed more TH-positive cells in SGZ-derivedNSCs and these SGZ-NSCs displayed a shorter differentiation time.Conclusion: SGZ-derived NSCs appear to be a more appropriate candidate for therapeutic purposes based onproliferation rate, neurosphere size, and Gfap and Nestin expression levels, as well as differentiation time and THexpression level after dopaminergic induction
Effects of steroid hormones on uterine tissue remodeling of mouse menopause model
Since the uterine is a sensitive tissue to steroid hormones, the aim of the present study was to investigate the effects of 17β-estradiol (E2) and progesterone (P4) alone or in combination on morphological and morphometrically parameters of ovariectomized mouse uterus.Adult virgin female mice (8-10 weeks old)were ovariectomized and treated with E2, P4, E2 followed by P4 and the oil vehicle alone for 5-days period. Uterine tissue was removed, and processed for histology assessment. The total uterine diameter were significantly higher (P < 0.05) following E2 treatment and Maximum diameter of uterine lumen, myometrium and endometrium were recorded after this treatment regimen. Sequential treatment with oestradiol and then progesterone caused both mitotic activity and cell degeneration. P4 treatment induced signs of active secretion in the endometrium glands and symptoms of degeneration and cell death. Estradiol treatment induced growth of uterine tissue. Subsequent treatment with progesterone stimulated uterine tissues to reach maximum size and maturity which is necessary to modify the uterus in preparation for pregnancy.
Intravenous Transplantation of Adipose-Derived Mesenchymal Stem Cells Promoted The Production of Dopaminergic Neurons and Improved Spatial Memory in A Rat Model of Parkinson’s Disease
Objective: Parkinson’s disease (PD) is a neurodegenerative disorder described by the dynamic decline of dopaminergicneurons in the substantia nigra pars compacta (SNpc). Stem cell transplantation is a new therapeutic strategy in thetreatment of PD. The objective of the study was to assess the impact of intravenous infusion of adipose-derivedmesenchymal stem cells (AD-MSCs) on memory disorder in Parkinsonian rats.Materials and Methods: In this experimental study, male Wistar rats were randomly divided to four groups containingsham, cell treatment, control, and lesion. The cell treatment group received intravenous injection of AD-MSCs 12 daysafter PD induction by bilateral injection of 6-hydroxydopamine. Four weeks after lesion formation, spatial memorywas examined using the Morris water maze (MWM) assessment. The rats’ brains were removed and assessed bybromodeoxyuridine (BrdU), tyrosine hydroxylase (TH), and glial fibrillary acidic protein (Gfap) immunostaining.Results: Statistical analyses revealed a significant addition and reduction in time spent and escape latency in the targetquadrant, respectively, in the cell group as compared to the lesion group. Also, BrdU-labeled cells were present in thesubstantia nigra (SN). The density of TH-positive cells was significantly increased in the AD-MSCs transplantation groupas compared to the lesion group, and the density of astrocytes significantly diminished in the AD-MSCs transplantationgroup as compared to the lesion group.Conclusion: It appears that AD-MSCs treatment for Parkinson’s could decrease the density of astrocytes and promotethe density of TH-positive neurons. It appears that AD-MSCs could improve spatial memory impairment in PD
Transdifferentiation of Bone Marrow Stromal Cells into Tyrosine Hydroxylase Immunoreactive Cells Associated with Angiogenesis in Parkinsonian Rats
Objective: This study is an attempt to examine the transdifferentiation of bone marrowstromal cells (BMSCs) into tyrosine hydroxylase immunoreactive cells in parkinsonian ratsassociated with angiogenesis.Materials and Methods: In this study, Sprague-Dawley rats received unilateral stereotaxicinjections of 6-hydroxydopamine(6-OHDA) into the left corpus striatum and then weredivided into two groups. One group, the negative control, received only medium while theother group was treated with BMSCs. BMSCs were harvested from femur bones, labeledwith bromodeoxyuridine (BrdU) and then transplanted into parkinsonian rats, where a behavioralstudy and immunohistochemistry were used to evaluate the treatment.Results: The results showed statistically significant improvement in rotational behavior.Anti-BrdU antibody showed engraftment of the transplanted cells at the transplantationsite. Additionally, double immunolabeling confirmed that these cells were positive for neurofilament-200 and tyrosine hydroxylase (TH).Conclusion: It may be concluded that BMSCs transplants could engraft and differentiateinto TH immunoreactive cells which may cause recovery from motor deficits. Also, BMSCsmay contribute to angiogenesis at the transplantation site
Spontaneous Expression of Neurotrophic Factors and TH, Nurr1, Nestin Genes in Long-term Culture of Bone Marrow Mesenchymal Stem Cells
Objective: It has been reported that rat bone marrow stromal cells (BMSCs) can be spontaneouslydifferentiated into neural-like cells without any supplemental growth factorsand/or chemical treatment after long-term culture.This study aims to determineWhether,growth factors secreted by MSCs could induce self-differentiation into neural-like cells ina long-term culture.Materials and Methods: This study consisted of two groups: i. rat BMSCs (passage 5)were cultured in alfa- minimal essential medium (α-MEM) and 10% fetal bovine serum(FBS) without the addition of inducer and exchanging medium for three weeks, as theexperimental group and ii.rat BMSCs (passage 5) as the control group. Each group wasanalysed by reverse transcriptase polymerase chain reaction (RT-PCR) to evaluate theexpressions of neurotrophic factors and neural marker genes.Statistical analyses were carried out using one-way analysis of variance (ANOVA) andTukey’s multiple comparison with SPSS software (version 16). P< 0.05 was consideredstatistically significant.Results: The experimental group (fifth passage of BMSCs) obtained from adult rats spontaneouslydifferentiated into neural precursor cells after long-term culture. Cultured cellsexpressed tyrosine hydroxylase (TH), Nurr1 and nestin genes. Furthermore, some growingcells in suspension became neurosphere-like. Self-differentiated rat MSCs (SDrMSCs)expressed significantly higher levels of NGF (0.96 ± 0.16), nestin (0.63 ± 0.08), and Nurr1(0.80 ± 0.10) genes (p<0.05).Conclusion: In this study, we reported that rMSCs in long-term culture underwent spontaneoustransformation to neural precursors without the supplement of growth factors andspecific chemicals. Cells expressed neural markers such as: TH, Nurr1, and nestin genes
Effects of dimethyl sulfoxide on expression of neurotrophic factors and dopaminergic genes by mesenchymal stem cells
Introduction: Neurotrophic factors play an important role in differentiation of mesenchymal stem cells (MSCs) into dopaminergic neurons. The objective of this work was to determine the expression of brain derived neurotrophic factor (BDNF), nerve growth factor (NGF), ciliary neurotrophic factor (CNTF), glial cell derived neurotrophic factor (GDNF), NT3 and NT5 and dopaminergic genes such as tyrosine hydroxylase in MSCs before and after dimethyl sulfoxide (DMSO) induction. The aim of this study was to evaluate the differentiation of MSCs induced by DMSO.Materials and Methods: MSCs obtained from the femur and tibia of adult rats were cultured in α-MEM mediums supplemented with serum. Fourth passage of MSCs were cultured in two different types of medium: 1- α-MEM containing 10% serum as control group (untreated cells) and 2- α-MEM containing 2% DMSO as treated cells. Identification and determination of the purity of MSCs were examined by immunocytochemistry staining. Reverse transcription polymerase chain reaction (RT-PCR) was used to study mentioned genes expression of the MSCs.Results: These results represented that MSCs were immunopositive for alkaline phosphatase, anti- fibronectine and anti- CD90. In addition the semiquantative study of PCR method showed that some neurotrophic factors (BDNF, NGF, CNTF, GDNF) expression in the untreated MSCs increased significantly compared with that of DMSO-treated group (P<0.05).Conclusion: MSCs play an important role in the treatment of Parkinson's disease by expression of neurotrophic factors and some dopaminergic gene
In Vitro Expression of BDNF GDNF NGF NT3 and NT4/5 Genes in Selegiline Induced Bone Marrow Stromal Cells
Objective: Two types of stem cells are found in the bone marrow: hematopoietic stemcells and marrow stromal cells (MSCs). Is it possible to induce the differentiation of bonemarrow stromal cells into neural cells in vitro and subsequently transplant them into thebrain? This might help repair neural lesions observed in some neurodegenerative disoders such as Parkinson’s disease (PD).Materials and Methods: In this study, cultured MSCs were incubated in serum free mediumcontaining 10-8 M selegiline for 24 hours and cells were cultured for another 48 hoursin α minimal essential medium (α-MEM) containing 20% fetal bovine serum (FBS).Then selegiline-treated cells were immunostained for neuronal markers such as NF-200and TH.Results: Cell counting results showed that Selegiline at doses of 10-6, 10-7 and 10-8 Mincreased the mean percent of viable cells. The most effective dose of Selegiline for diferentiationof bone marrow stromal cells (BMSCs) was 10-8 M. Molecular studies indcatedthat the expression of BDNF, GDNF, NGF, NT3, and NT4/5 genes were increased inSelegiline-treated cells compared to non-treated group.Conclusion: BMSCs can be directed to a neural fate in vitro and can be considered as acell source in neurological disorders for autograft therapy
The study of neurotrophic factor genes expression of human adipose stem cells cultured in serum-containing and serum-free media
Background: Fetal bovine serum (FBS) is immunogenic for human and may transmit infection in the case of transplantation. So, this study aimed to compare the proliferation and survival rates of human adipose stem cells (hASCs), and their neurotropic factor genes expression in serum-containing and serum-free media.
Materials and Methods: In this experimental study, stem cells were extracted from the abdominal subcutaneous adipose tissue of 15 cesarean women and cultured in α-MEM containing 10% of FBS or serum-free medium. The stemness of fourth passage of the cells was confirmed using the flow cytometry method, and their differentiation into adipocytes and osteocytes was also confirmed. Cell proliferation and survival were assessed using hemocytometry and MTT [3- (4,5-Dimethyltiazol-2-yl) -2,5-Diphenyltetrazolium bromide] methods, respectively. In addition, the expression of neurotrophic factor genes was analyzed by the real-time polymerase chain reaction method.
Results: The cells had positive response to CD44, CD73, CD90, and CD105 markers, while they responded negatively to CD34 and CD45 markers and had the ability to differentiate into adipocytes and osteocytes. The survival and proliferation of the cells cultured in the serum-based medium for 48 hours were significantly increased compared to those cultured in the serum-free medium. Moreover, serum resulted in a significant increase in BDNF and NT-3 genes expression, compared to the cells cultured in the serum-free medium.
Conclusions: More suitable cells can be provided for transplantation with serum deletion and culture medium optimization. The results can be matched to find an appropriate replacement for FBS
Effect of exogenous estrogen treatment on hippocampal neurogenesis in ovariectomized mice
Background and Objective: Adult neurogenesis occurs in most mammalian species in two main areas of brain: 1- subventricular zone 2- the dentate gyrus of the hippocampus. Many factors such as 17-B estradiol affect neurogenesis in the hippocampus. The aim of this study was to investigate the effect of exogenous 17-B estradiol on neurogenesis and astrocyte functions in the ovariectomized mice.
Methods: In this experimental study; NMRI mice were allocated into five experimental groups including Sham, Control, Treatment with single dose of 17-B estradiol two weeks after ovariectomy (OVX) and were sacrificed 24 hours later, Treatment with single dose of 17-B estradiol two weeks after Ovx and were sacrificed 48 hours later and  Treatment with single dose of Seasame Oil 2 weeks after OVX and were sacrificed after 24 hours. Animals were transcardially perfused with paraformaldehyde. Brains were removed and its sections for cresyl fast violet staining and GFAP immunohistochemistry were prepared. Cells were counted and investigated.
Results: Neuronal density and Proliferation of hippocampal progenitor cells in the CA1 region of 17-B estradiol treated mice significantly increased up to 24 hours (P<0.05). Density of glia and particularly astrocytes in different regions of the hippocampus significantly reduced after treatment with 17-B estradiol (P<0.05).
Conclusion: Density of hippocampal CA1 neurons are influenced by 17-B estradiol. Also, density and morphology of glia cells, especially astrocytes in different regions of the hippocampus are affected by 17-B estradiol