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عضویت

جستجوی مقالات مرتبط با کلیدواژه "bone marrow mesenchymal stem cells" در نشریات گروه "پزشکی"

  • Ruben Chailakhyan, Alla Grosheva, Nataliya Vorobieva, Vladimir Yusupov, Alexander Sviridov *
    Introduction

     The purpose of this study is to achieve a significant increase in the proliferative activity of mesenchymal stem cells (MSCs) of the bone marrow (BM) at early passages after laser exposure to a suspension of these cells and to estimate the effect of light and heat components of laser radiation on the proliferation of BM MSCs.

    Methods

     The studies were performed on rats with BM MSCs. MSC suspension was placed into the wells and heated by using laser radiation (980 nm wavelength) or a water bath at 70 °C providing similar temperature dynamics. The studies were carried out in 3 comparison groups: (1) control suspension of MSCs, which was not subjected to heating in a water bath or laser exposure; (2) MSC suspension, which was heated in a water bath; and (3) suspension of MSCs, which was subjected to laser exposure. The exposure times for the 2nd and 3rd experimental groups were 10- 50 seconds.

    Results

     Under optimal parameters of laser action on the suspension of BM MSCs, a six-fold increase in the number of BM MSCs colonies was registered compared to the control. The role of the light and heat components of laser exposure to MSCs was determined by comparable heating of a suspension of BM MSCs in a water bath, at which only a twofold increase in the number of colonies was maximally obtained.

    Conclusion

     The increase in the MSC proliferation activity occurs due to their ThermoPhotobiomodulation. The result obtained is important for practical use in cell transplantation in the treatment of traumatic injuries of bone, cartilage, and tendon tissues when a rapid and multiple increase in the initial number of autologous BM MSCs is required.

    Keywords: Bone Marrow Mesenchymal Stem Cells, Laser Heating, Thermo-Photobiomodulation, Cell Therapy
  • مقدمه

    سیکلوفسفامید (CP) اثرات منفی بر سیستم تولید مثل دارد. سلول های بنیادی و متابولیت های آنها جهت افزایش باروری پس از شیمی درمانی مورد استفاده قرار گرفته اند.

    هدف

    این مطالعه با هدف بررسی تاثیر محیط مشروط (Conditioned medium: CM) مشتق شده از سلول های بنیادی مزانشیمی مشتق از مغز استخوان بر اثرات سمی CP بر بیضه ها انجام شد.

    مواد و روش ها

    سلول های بنیادی مزانشیمی مغز استخوان استخراج، محیط مشروط جمع آوری و 25 برابر تغلیظ شد. 24 سر موش های صحرایی نر نژاد ویستار (8 هفته، 250-200 گرم) به صورت تصادفی به 4 گروه تقسیم شدند: کنترل، CP، CP+DMEM، CP+CM .CP به میزان 100 میلی گرم بر کیلوگرم به شکل تک دوز تزریق شد. دو هفته پس از تجویز سیکلوفسفامید، CM از طریق مجرای وابران بیضه تزریق شد. پارامترهای اسپرم، هیستوپاتولوژی بیضه و سطح تستوسترون دو ماه پس از درمان بررسی شد. بیان ژن های Bax و Bcl2 توسط real-time polymerase chain reaction مورد ارزیابی قرار گرفت.

    نتایج

    CP بر بافت شناسی بیضه (001/0 < p) و کیفیت اسپرم (001/0 < p) تاثیر منفی داشت. CP بیان ژن های مرتبط با آپوپتوز را تغییر داد (001/0 < p). درمان با CM بیان Bax را صورت معنی داری کاهش داد (001/0 < p)، در حالی که بیان Bcl2 را به طور معنی داری افزایش نشان داد (01/0 = p). CP تعداد اسپرم (03/0 = p)، زنده مانی (001/0 < p) تحرک (001/0 < p)، تعداد اسپرماتوگونی (001/0 < p) و ضخامت اپیتلیال لوله های بیضه (02/0 = p) را بهبود داد.

    نتیجه گیری

    این یافته ها نشان می دهد که CM تولید شده از سلول های بنیادی مزانشیمی مشتق از مغز استخوان ممکن است در درمان های ناباروری ارزشمند باشد و عوارض جانبی CP را کاهش دهد.

    کلید واژگان: سلول های بنیادی مزانشیمی مغز استخوان, سیکلوفسفامید, محیط شرطی شده, آپوپتوز, اسپرماتوژنز
    Zeynab Esmailpour, Soheila Madadi, Maryam Baazm *
    Background

    Cyclophosphamide (CP) has some negative effects on the reproductive system. Stem cells and their metabolites are being utilized to enhance fertility after chemotherapy.

    Objective

    This study aimed to investigate the impact of conditioned medium (CM) derived from bone marrow mesenchymal stromal stem cells (BM-MSCs) on the toxic effects of CP on testicles.

    Materials and Methods

    BM-MSCs were isolated, a CM was collected and 25-fold concentrated. 24 male Wistar rats (8 wk, 200-250 gr) were randomly divided into following groups: control, CP, CP+Dulbecco’s Modified Eagle Medium (DMEM), CP+CM. CP was given at a single dose of 100 mg/kg. 2 wk after the CP administration, CM was injected into the testicular efferent duct. Sperm parameters, testicular histopathology, and the level of testosterone were analyzed 2 months after treatment. The expression of B-cell lymphoma 2 (Bcl2) and Bcl2-associated X protein (Bax) genes were evaluated by real-time polymerase chain reaction.

    Results

    CP had a negative effect on testis histology (p < 0.001) and sperm quality (p < 0.001). It changed the expression of genes associated with apoptosis (p < 0.001). Treatment with CM reduced the expression of Bax (p < 0.001), while significantly increasing the expression of Bcl2 (p = 0.01). It improved sperm count (p = 0.03), viability (p < 0.001), motility (p < 0.001), spermatogonial count (p < 0.001), and epithelial thickness of testicular tubules (p = 0.02).

    Conclusion

    These findings suggest that CM produced from BM-MSCs may be valuable for therapeutic approaches in reproductive medicine and may lessen the side effects of CP.

    Keywords: Bone marrow mesenchymal stem cells, Cyclophosphamide, Conditioned medium, Apoptosis, Spermatogenesis
  • MohammadSadegh Gholami Farashah, Maryam Javadi, Jafar Soleimani Rad, Seyed Kazem Shakouri, Solmaz Asnaashari, Siavoush Dastmalchi, Sadeneh Nikzad, Leila Roshangar*
    Purpose

     Exosomes are natural nanoparticles that participate in intercellular communication through molecular transport. Recently, due to their membrane vesicular structure and surface proteins, exosomes have been used extensively in the research field of drug delivery. Osteoporosis is an inflammation in which the cellular balance of bone tissue is disturbed that reduces bone density and making bone prone to abnormal fractures with small amount of force. Utilizing estrogen is one of the main therapeutic strategies for osteoporosis. Despite the positive effects of estrogen on bone tissue, changes in the natural estrogen levels of the body can cause a number of diseases such as different types of cancer. Therefore, designing a therapeutic system which controls more accurate tissue targeting of estrogen seems to be a rational and promising practical approach.

    Methods

     In this study, bone marrow mesenchymal stem cells (BMMSCs)-derived exosomes were loaded by estradiol using two different methods of drug loading, namely incubation and sonication methods and then the survival effects of the drug loaded exosomes on BMMSCs was investigated.

    Results

     Examination of size, shape, and surface factors of exosomes in different states (pure exosomes and drug-loaded exosomes) showed that the round morphology of exosomes was preserved in all conditions. However, the particles size increased significantly when loaded by sonication method. The increased survival of BMMSCs was noted with estradiol-loaded exosomes when compared to the control group.

    Conclusion

     The results suggest that estradiol-loaded exosomes have potential to be used as nano-drug carriers in the treatment of osteoporosis.

    Keywords: Osteoporosis, Exosome, Drug delivery, 17β-estradiol, Bone marrow mesenchymal stem cells
  • Saeed Rahimi, Mahnaz Ahrabi, Mohammad Samiei, Leila Roshangar, Behnaz Ahrabi, Behnam Hashemi, Shahriar Shahi, Naghmeh Rahimi Darehchi
    Introduction

    Exposure to pulsed electromagnetic field (PEMF) has been revealed to affect the differentiation and proliferation of human mesenchymal stem cells derived from dental pulp multipotent stromal stem cells (DP -MSCs). This study aimed to investigate the differentiation effect of electromagnetic fields (EMFs) on the DP-MSC.

    Materials and Methods

    PEMF was produced by a system comprising a multi -meter autotransformer, solenoid coils, and teslameter. This study included 10 groups of DP-MSCs which underwent different electromagnetic radiation time and beam intensity. Three samples tested for each group. The effect of PEMF with the intensity of 0.5 and 1 mT (mili Tesla) and 50 Hz on the proliferation rate of DP-MSC was evaluated at 20 and 40 minutes per day for seven days. MTT assay was applied to determine the growth and proliferation of DP-MSC. Gene expression of DMP1 for differentia tion of DPSCs to odontoblasts was confirmed by Real Time PCR., ANOVA statistical analysis and Kruskal-Wallis test were used to analyze the data.

    Results

    The survival in all exposure groups was significantly higher than that in control except in the group of 40 minutes, 1 mT (P<0.05). In 20 minutes, 0.5 mT exposure, the survival intensity is significantly more than others (P<0.05). In general, the intensity of survival was recorded, 20, 0.5 mT ≥20, 1 mT ≥40, 0.5 mT≥40, 1 mT respectively. Therefore, according to the obtained results, ELF-EMF increases the survival of cells except for one case (40 minutes, 1mT), even though the effective underlying mechanisms in this process are still unclear.

    Conclusions

    The results obtained promise that in the future, by placing an important part of the pulp next to the electromagnetic field, the lost part of the pulp can be reconstructed and the dentin barrier can be created.

    Keywords: Bone Marrow Mesenchymal Stem Cells, Dental Pulp, Dental Pulp -derived Mesenchymal Stem Cells, Pulsed Electromagnetic Field
  • Li Yan, Yao Xiu, Lin Yanjun, Xing Yifeng, Liu Chaowei, Xu Jianghan, Wu Dong *
    Objective (s)

    Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is an essential stage in bone formation. Autophagy plays a pivotal role in the self-renewal potential and pluripotency of stem cells. This study aimed to explore the function of autophagy-related genes during osteogenic differentiation of BMSCs.

    Materials and Methods

    The differentially expressed autophagy-related genes (ARGs) were obtained from the GEO and HADb databases. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using R software. The PPI and hub gene mining networks were constructed using the STRING database and Cytoscape. Finally, the RT-qPCR was conducted to validate the expression level of ARGs in BMSCs.

    Results

    Thirty-seven differentially expressed ARGs were finally obtained, including 12 upregulated and 25 downregulated genes. GO and KEGG enrichment analysis showed that most of these genes were enriched in apoptosis and autophagy. The PPI network revealed strong interactions between differentially expressed ARGs. The expression level of differentially expressed ARGs tested by RT-qPCR showed 6 upregulated ARGs, including FOXO1, MAP1LC3C, CTSB, FOXO3, CALCOCO2, FKBP1A, and 4 downregulated  ARGs, including MAPK8IP1, NRG1, VEGFA, and ITGA6 were consistent with the expression of high-throughput sequencing data.

    Conclusion

    We identified 37 ARGs during osteogenic differentiation using bioinformatics analysis. FOXO1, MAP1LC3C, CTSB, FOXO3, CALCOCO2, FKBP1A, MAPK8IP1, NRG1, VEGFA, and ITGA6 may regulate osteogenic differentiation of hBMSCs by involving autophagy pathway. This study provides new insight into the osteogenic differentiation of hBMSCs and may be available in developing therapeutic strategies for maxillofacial bone defects.

    Keywords: Autophagy, Bioinformatics, Bone marrow mesenchymal stem cells, bone regeneration, Osteogenesis
  • علیرضا شمس، عاطفه شموسی*
    مقدمه و هدف

    تمایز سلولهای بنیادین مزانشیمی مغز استخوان  BMMSC (bone marrow mesenchymal stem cell) به رده های مختلف سلولی بررسی شده است. سیگنال های مختلف بیوشیمیایی و توپوگرافیک می توانند در تکثیر و تمایز سلولهای بنیادین به یک رده سلولی خاص تاثیر بگذارند. هدف از این مطالعه ارزیابی توانایی BMMSC برای تمایز به سلولهای شبه نورونی است.

    مواد و روش ها

    از داربست ریخته گری انجمادی و خشک انجمادی  ژلاتین/ پلی لاکتیک-کو-گلیکولیک اسید (PLGA) با نانوذرات آهن (MNPs) و بدون نانوذرات آهن (MNPs-free)  به عنوان یک ساختار توپوگرافی سه بعدی استفاده شد. تمایز BMMSC  به سلولهای شبه نورون تحت اثر محیط های نوروژنیک به مدت 20 روز انجام شد. میزان تکثیر BMMSC  با تست MTT  و بیان مارکرهای خاص نورون با روش ایمونوهیستوشیمی بررسی شد.  در این مطالعه،  روش آماری آنالیز واریانس یک طرفه ANOVA، Post Hock Tukey و آزمون نان پارامتریک Mann-Whitney استفاده شد.

    نتایج

    داربست ریخته گری انجمادی  ژلاتین-PLGA- NPs حاوی نانوذرات آهن ساختار مناسبی برای تمایز به سلولهای شبه نورونی فراهم می کند. نتایج بقا سلولی (میانگین 7600/0 و انحراف معیار 02000/0 بعد از 72 ساعت) و بیان ژنها (میانگین 00/79 و انحراف معیار 000/1) نشان می دهد که میزان تکثیر سلول ها بر روی داربستهای فریز کست شده حاوی نانوذرات بیشتر از داربستهای فریزداری شده می باشد (001/0>P).

    نتیجه گیری

    داربست ریخته گری انجمادی ژلاتین-PLGA حاوی نانوذرات آهن می تواند انتخاب مناسبی برای ترمیم و بازسازی سیستم عصبی باشد.

    کلید واژگان: تمایز, سلولهای بنیادی مغز استخوان, داربست, ریخته گری انجمادی, خشک انجمادی
    Alireza Shams, Atefeh Shamosi *
    Background and Objective

    Differentiation of bone marrow mesenchymal stem cells (BMMSC) has studied in different cell lines. Variety of biochemical and topographic signaling could influence and change cells differentiation and proliferation of a special cell line. The aim of this study was to evaluate potential differentiating of BMMSC to neuronal-like cells.

    Materials and Methods

    Freezed casting and freezed drying scaffolds of gelatin/polylactic-co-glycolic acid (PLGA) with iron nanoparticles (MNPs) and without iron nanoparticles (MNPs-free) was used as a three-dimensional topographic structure. BMMSC differentiation to neuronal-like cells by neurogenic culture was done in 20 days. BMMSC proliferation was studied by MTT assay. Gene expression of specific neuronal markers was studied by immunohistochemistry methods. In this study, one-way ANOVA, post hoc Tukey and Mann-Whitney nonparametric tests were used. 

    Results

    Gelatin PLGA- MNP freezed casting scaffold containing Fe nanoparticles provides a suitable structure for differentiation into neuron-like cells. The results of cell survival (Mean 0.7600 and standard deviation 0.02000 after 72 hours) and gene expression (Mean 79.00 and standard deviation 1.000) showed that the rate of cell proliferation on freezed casting scaffolds containing nanoparticles is higher than that of freezed drying scaffolds (P<0.001).

    Conclusion

    Freezed casting gelatin PLGA-MNP scaffold could be an appropriate choice for repair and regeneration of nervous system.

    Keywords: Bone marrow mesenchymal stem cells, Differentiation, Freezed casting, Freezed drying
  • Mohammad Hussein Abnosi*, Setarehsadat Hosseini
    Background

    Cell toxicity due to diethanolamine (DEA) is well known but no data are available regarding its mechanism. The present study investigated the cell viability and proliferation ability of rat bone marrow mesenchymal stem cells (BMSCs) treated with DEA.

    Methods

    At 3rd passages, BMSCs were treated for 12, 24, and 48 hours with 0.025 to 16 mM of bis(2- ethylhexyl) phthalate. The cell viability was estimated using the trypan blue and MTT, then 1, 4, and 16 mM, and 48 hours were selected as well. Next, other parameters were determined, including proliferation ability, cell morphology, sodium and potassium levels, as well as the concentration of calcium, total protein, and the activity of metabolic enzymes (i.e., alanine transaminase [ALT], aspartate transaminase [AST], and lactate dehydrogenase [LDH]). Finally, malondialdehyde (MDA), total antioxidant capacity (TAC), and the activity of antioxidant enzymes such as superoxide dismutase and catalase were measured based on the aim of the study.

    Results

    Based on the results, the viability reduced significantly from 0.6 mM at 12 hours and 0.2 for 24 and 48 hours (P<0.05). In addition, the proliferation ability showed a significant reduction (P<0.05) while the activity of LDH, ALT, and AST increased significantly (P<0.05). The level of electrolytes at 1 mM treatment demonstrated no change (P>0.05) whereas 4 mM concentration caused a decline in the calcium level while increased the sodium significantly (P<0.05). The results further revealed that the level of MDA increased although the activity of antioxidant enzymes and TAC represented a significant reduction (P<0.05). No change was detected in the morphology of nuclei while cytoplasm shrinkage and roundness were observable.

    Conclusion

    In general, the findings of this study showed that DEA reduced the viability and proliferation of BMSCs via metabolic change, electrolyte imbalance, and the induction of oxidative stress.

    Keywords: Bone marrow mesenchymal stem cells, Di-ethanolamine, Cell viability, Cell proliferation, Oxidative stress
  • Saba Behzadi Fard, Zohreh Mazaheri*, Nasim Ghorbanmehr, Mansoureh Movahedin, Mahin Behzadi Fard, Mohammad Ali Gholampour

    In vitro derivation of germ cells from different stem cells sources has been challenging in the treatment of male infertility. MicroRNAs (miRNAs) have an essential role in gene expression at post-transcriptional level. The aim of this research was to find more about miRNA-17 and miRNA-146 expression during differentiation of spermatogonial stem cell like cells (SSC like cells) from mouse bone marrow mesenchymal stem cells (BMSCs) through bone morphogenic protein 4 (BMP4) and retinoic acid (RA) induction. BMSCs were treated with BMP4 to produce primordial germ cell like cells (PGC like cells). The cells were differentiated into SSC like cells by an inducer cocktail including RA, leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF). The PGC like cells and SSC like cells were evaluated for pluripotency (Nanog, Oct-4) and germ cell specific gene (Piwil2, Plzf, Dazl, and Stra8) expression, protein expression (Plzf, Stra8), and miRNA-17 and miRNA-146 mRNA expression. Our results showed that BMP4 lead to Dazl up-regulation and Nanog down-regulation expression in PGC like cells. RA up-regulated Stra8 and Piwil2, and down-regulated Nanog and Oct-4. MiRNA-17 and miRNA-146 expression decreased significantly in SSC like cells after RA treatment. This research indicated the aberrant miRNA-17 and miRNA-146 expression in SSC like cells in comparison with SSCs. Down-regulation of the two miRNAs by using RA in the stimulated undifferentiated state could be probably one of the key factors of SSC like cells arrest.

    Keywords: Bone marrow mesenchymal stem cells, spermatogonial stem like cells, differentiation, miRNA-17, miRNA-146
  • Fereshteh Dorazehi, Mohammad Nabiuni *, Hanieh Jalali
    Scaffolds derived from decellularized tissues provide a natural microenvironment for cell culture. Embryonic cerebrospinal fluid (e-CSF) contains factors which play vital roles in development of the nervous system. This research was aimed to survey the effect of Wistar rat e-CSF on neural differentiation of bone marrow derived mesenchymal stem cells (BM-MSCs) cultured on the human amniotic membrane (AM). BM-MSCs were collected from femurs and tibias, and were cultured in Dulbecco's Modified Eagle's Medium. The placenta was harvested from healthy women during cesarean section and AM was acellularized using EDTA and physical scrubbing. e-CSF was harvested from rat fetuses at E17. Adequate numbers of BM-MSCs were cultured on acellularized membrane, and were treated with E17 CSF for 7 days. MTT (3-(4, 5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide) assay confirmed the survival and proliferation of BM-MSCs cultured on AM derived scaffold. Hematoxylin/eosin staining and scanning electron microscopy showed the morphological and the structural changes of BM-MSCs throughout the culture and treatment with e-CSF. The results of immunocytochemistry showed that microtubule associated protein 2 and beta-III tubulin were expressed in BM-MSCs cultured on acellular amnion scaffold and treated with e-CSF. Our results showed for the first time that the combination of acellular AM as a natural scaffold and e-CSF as a source of neurological factors could effectively improve the BM-MSCs cultivation and differentiation.
    Keywords: Cerebrospinal Fluid, Bone Marrow Mesenchymal Stem Cells, Neural Differentiation, Amnion Membrane Derived Scaffold
  • Ali Rafat, Amaneh Mohammadi Roushandeh, Akram Alizadeh, Nasrin Hashemi-Firouzi, Zoleikha Golipoor *
    Objective
    Mesenchymal stem cells (MSC) from various sources have the potentials to positively affect regenerative medicine. Furthermore, pre-conditioning strategies with different agents could improve the efficacy of cell therapy. This study compares the effects of an anti-inflammatory and antioxidant agent, melatonin, on protection of bone marrow-derived MSCs (BMSCs) and adipose tissue-derived MSCs (ADSCs).
    Materials And Methods
    In this experimental study, rat BMSCs and ADSCs were isolated and expanded. Pre-conditioning was performed with 5 µM melatonin for 24 hours. Cell proliferation and viability were detected by MTT assay. Expression of BAX, BCL2, melatonin receptors and osteocalcin genes were evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR). Also, apoptosis was detected with tunnel assay. Osteogenic differentiation was analyzed using alizarin red staining.
    Results
    No significant increase was found in cell viability between BMSCs and ADSCs after melatonin preconditioning. Following melatonin preconditioning, BAX expression was significantly down-regulated in both ADSCs and BMSCs (P
    Conclusion
    Here we have shown that the effects of preconditioning on melatonin expression in ADSCs are higher than those in BMSCs. These findings could be used in adoption of a proper preconditioning protocol based on the sources of MSCs in specific clinical applications, especially in bone regeneration.
    Keywords: Apoptosis, Bone Marrow Mesenchymal Stem Cells, Melatonin, Osteogenesis
  • Huichao Wang, Chunbo Li, Jianming Li, Yingjie Zhu, Yudong Jia, Ying Zhang, Xiaodong Zhang, Wenlong Li, Lei Cui, Wuyin Li, Youwen Liu *
    Objective(s)
    Naringin has been reported to regulate bone metabolism. However, its effect on osteogenesis remains unclear. The aim was to investigate the effect of naringin on osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) through the activation of the ERK signaling pathway in osteogenic differentiation.
    Materials And Methods
    Annexin V-FITC assay and MTT assay were used to measure the effect of naringin on cytotoxicity and proliferation of hBMSCs, respectively. Alkaline phosphatase activity analysis, Alizarin Red S staining, Western blotting, and real-time PCR assay were used to evaluate both the potential effect of naringin on osteogenic differentiation and the role of ERK signaling pathway in osteogenic differentiation.
    Results
    Our results showed that naringin had no obvious toxicity on hBMSCs, and could significantly promote the proliferation of hBMSCs. Naringin also enhanced the osteogenic differentiation of hBMSCs and increased the protein and mRNA expression levels of osteogenic markers such as Runx-2, OXS, OCN, and Col1 in a dose-dependent manner. In addition, we found that the enhancing effect of naringin on osteogenic differentiation was related to the activation of phosphor-ERK, with an increase in duration of activity from 30 min to 120 min. More importantly, both the enhancing effect of naringin on osteogenic differentiation and the activity effect of naringin on ERK signaling pathway were reversed by U0126 addition.
    Conclusion
    Our findings demonstrated that naringin promoted proliferation and osteogenesis of hBMSCs by activating the ERK signaling pathway and it might be a potential therapeutic agent for treating or preventing osteoporosis.
    Keywords: Bone marrow-mesenchymal stem cells, ERK signaling pathway, Naringin, Osteogenic differentiation
  • Iman Razeghian Jahromi, Davood Mehrabani, Ali Mohammadi, Mohammad Mahdi Ghahramani Seno, Mehdi Dianatpour, Shahrokh Zare, Amin Tamadon
    Objective(s)
    Nowadays much effort is being invested in order to diagnose the mechanisms involved in neural differentiation. By clarifying this, making desired neural cells in vitro and applying them into diverse neurological disorders suffered from neural cell malfunctions could be a feasible choice. Thus, the present study assessed the capability of fetal brain extract (FBE) to induce rat bone marrow-derived mesenchymal stem cells (BM-MSCs) toward neural cells.
    Materials And Methods
    For this purpose, BM-MSCs were collected from rats and cultured and their mesenchymal properties were confirmed. After exposure of the BM-MSCs to fetal brain extract, the cells were evaluated and harvested at days 3 and 7 after treatment.
    Results
    The BM-MSCs that were exposed to FBE changed their appearance dramatically from spindle shape to cells with dendrite-like processes. Those neural like processes were absent in the control group. In addition, a neural specific marker, vimentin, was expressed significantly in the treatment group but not in the negative control group.
    Conclusion
    This study presented the FBE as a natural neural differentiation agent, which probably has required factors for making neurons. In addition, vimentin overexpression was observed in the treated group which confirms neuron-like cell differentiation of BM-MSCs after induction.
    Keywords: Bone marrow-mesenchymal-stem cells, Differentiation, Fetal brain extract, Neural cell, Vimentin
  • Mahdieh Hajian Monfared, Bagher Minaee, Tayebeh Rastegar, Ebrahim Khrazinejad, Mohammad Barbarestani *
    Objective(s)
    Although many researchers have confirmed induction of germ cells from bone marrow mesenchymal stem cells (BMMSCs), there are no reports that confirm spontaneous differentiation of germ cells from BMMSCs. In this study, we have evaluated the effect of adult Sertoli cell condition medium (SCCM) as a mutative factor in the induction of germ cells from BMMSCs.
    Materials And Methods
    BMMSCs were collected from the bone marrow of 6-8-week old NMRI mice and their mesenchymal entities were proven using superficial markers (expression of CD44 and CD73 and non-expresion of CD45 and CD11b) by fow cytometry. Their multi-potential entities were proved with differentiation to osteogenic and adipogenic cells for 21 days. Also isolated Sertoli cells were enriched using lectin coated plates and Sertoli cell condition medium (SCCM) was collected. Sertoli cells were identified by immunocytochemistry and Vimentin marker. The cells were then differentiated into germ cells with SCCM for 2 weeks. Finally induced cells were evaluated by RT-PCR and immunocytochemistry.
    Results
    Differentiation of mesenchymal stem cells to osteoblast and adipocyte showed their multi-potential property. Expression of CD44 and CD73 and non-expression of CD45 and CD11b confirmed mesenchyme cells. Immunocytochemistry and RT-PCR results showed expression of germ cells specific marker (Mvh).
    Conclusion
    This study confirmed the effect of SCCM as a motivational factor that can used for differentiation of germ cells from BMMSCs.
    Keywords: Bone marrow mesenchymal stem cells, Differentiation, Germ cells, Sertoli cell condition medium
  • سهیلا مددی درگاهی، مینا افتخارزاده، احمد مهدی پور، منصوره سلیمانی، مهدی مهدی زاده*
    زمینه و هدف
    امروزه به دنبال تایید وجود نوروژنز در مغز پستانداران بالغ، استفاده از سلول های بنیادی به عنوان یک روش درمانی مناسب برای بهبود بسیاری از بیماری های سیستم عصبی مورد توجه پژوهشگران قرار گرفته است. بدین ترتیب که با پیوند سلول های بنیادی، بازسازی نورونی در نواحی تخریب شده ایجاد می گردد. این مطالعه به منظور تعیین اثر پیوند سلول های بنیادی مزانشیمی مغز استخوان بر آسیب های وارده به هیپوکامپ انجام شد.
    روش بررسی
    در این مطالعه تجربی 28 سر موش صحرایی نر نژاد ویستار در چهار گروه 7 تایی کنترل، مدل، شاهد و درمان قرار گرفتند. حیوانات نوروتوکسین تری متیلین کلراید را به میزان 8 میلی گرم بر کیلوگرم وزن بدن از طریق تزریق داخل صفاقی دریافت کردند. یک هفته پس از دریافت نوروتوکسین، سلول های بنیادی به روش استریوتاکسی تزریق شد. شش هفته پس از تزریق سلول ها، حافظه فضایی موش ها به روش ماز آبی موریس بررسی شد. همچنین مطالعه بافتی با روش رنگ آمیزی نیسل و شمارش سلول های سالم توسط نرم افزار Olysia bio report انجام گردید.
    یافته ها
    به دنبال پیوند سلول های بنیادی مزانشیمی مغز استخوان، تعداد نورون های سالم در گروه درمان (15.19±74) در مقایسه با گروه شاهد (12.971±44.67) و گروه مدل (18.105±48.56) بیشتر بود (P<0.05). همچنین در آزمون ماز آبی موریس، به دنبال پیوند سلول های بنیادی مزانشیمی مغز استخوان، گروه درمان (189.18±378.35)، (13.67±31.30) مسافت و زمان کمتری برای رسیدن به سکوی مخفی طی نمود؛ ولی این کاهش نسبت به گروه شاهد (192.56±438.18)، (14.89±40.14) و گروه مدل (225.44±407.98)، (17.15±37.68) معنی دار نبود. همچنین مسافت طی شده در ربع هدف توسط گروه درمان (125.91±799.8) نسبت به گروه مدل (136.94±588.51) و گروه شاهد (86.47±546.48) افزایش آماری معنی داری یافت (P<0.05).
    نتیجه گیری
    استفاده از سلول های بنیادی مزانشیمی مغز استخوان باعث کاهش آسیب های وارده به هیپوکامپ به صورت افزایش تعداد نورون های پیرامیدال و بهبود حافظه گردید.
    کلید واژگان: نوروژنز, سلول های بنیادی مزانشیمی مغز استخوان, هیپوکامپ, تری متیلین کلراید, حافظه فضایی
    Madadi Dargahi S., Eftekharzadeh M., Mahdipour A., Soleimani M., Mehdizadeh M. *
    Background And Objective
    Stem cells are a suitable treatment method for improvement of central nervous system diseases. Neuron regeneration is occure in damaged region using stem cell transplantation. This study was done to determine the effect of bone marrow mesenchymal stem cells on memory and neuronal cells graft number in the trimethyltin chloride damaged hippocampus.
    Methods
    In this experimental study, 28 wistar male rats were allocated into four groups including control, model, Vehicle and treatment groups. Animals were received 8 mg/kg/bw of neurotoxin trimethyltin chloride by the intraperitoneal injection for causing damaged in hippocampus. One week after intraperitoneal injection of trimethyltin chloride, stem cells was injected by stereotaxy method. Six weeks after stem cells injection, the spatial memory was assessed by Morris water maze and histological studies were done by Nissl staining and normal cells count by Olysia bio report software.
    Results
    After bone marrow mesenchymal stem cells graft, the number of normal cells were more in the treatment group (74±15.190) in compared to the Vehicle (44.67±12.971) and Model (48.56±18.105) groups (P<0.05). Also in Morris water maze test, the treatment group (387.35±189.18), (31.30±13.67) spent shorter distance and escape latency to reach the hidden platform, but this reduced non significantly in compared to Vehicle (438.18±192.56), (40.14±14.89) and model (407.98±225.44), (37.68±17.15) groups. The model and Vehicle groups spent longer distance to reach the hidden platform in comparision with the control (275.45±165.10) group (P<0.05). Also the traveled distance in target quarter had significant increased in the treatment groups (799.80±125.91) in compared to model (588.51±136.94) and Vehicle (546.48±86.47) groups (P<0.05).
    Conclusion
    Using the bone marrow mesenchymal stem cells leads to reduce hippocampal lesions and increase the number of pyramidal neurons and improving memory in damaged hippocampus in animal model.
    Keywords: Neurogenesis, Bone marrow mesenchymal stem cells, Hippocampus, Trimethyltin chloride, Spatial memory
  • زهرا حاتم نیا*، فریبا اسماعیلی، فریبا هوشمند، لیلا ده دهی
    زمینه و هدف

    با دیابت نوع یک در نتیجه ی تخریب خود ایمنی سلول های بتای جزایر پانکراس ایجاد می شود. مطالعات اخیر نشان می دهد بسیاری از انواع سلول های بنیادی می توانند به عنوان منابع احتمالی برای به دست آوردن سلول های قابل پیوند تولید کننده انسولین (IPCs) در نظر گرفته شوند. در این مطالعه تمایز سلول های بنیادی مزانشیمی مغز استخوان به سلول های مولد انسولین با استفاده از عصاره پانکراس موش مورد بررسی قرار گرفت.

    روش بررسی

    در این مطالعه تجربی آزمایشگاهی از سلول های بنیادی مزانشیمی مغز استخوان موش کوچک آزمایشگاهی برای تمایز به سلول های انسولین ساز استفاده شد. سلول های تمایز یافته با استفاده از رنگ اختصاصی دیتیزون و آنتی بادی های ضد انسولین- پروانسولین و ضد رسپتور بتای انسولین مورد ارزیابی قرار گرفتند. همچنین بیان ژن اختصاصی سلول های پانکراسی یعنی pdx-I در این سلول ها با روش RT-PCR بررسی شد.

    یافته ها

    سلول های تمایز یافته مشتق از روش مستقیم مورفولوژی مشابه با سلول های بتای پانکراس نشان دادند. سلول های دیتیزون مثبت به صورت دستجات قرمز ارغوانی دیده شدند. نتایج بررسی RT-PCR بیان ژن اختصاصی سلول های بتا (pdx-I) را در سلول های تمایز مستقیم نشان داد. ایمنوفلورسنس وجود نشانگرهای اختصاصی سلول های بتا را دراین سلول ها به اثبات رساند.

    نتیجه گیری

    نتایج این مطالعه نشان داد که سلول های بنیادی مزانشیمی مغز استخوان می توانند در حضور عصاره پانکراس به سلول های مولد انسولین تمایز یابند؛ لذا استفاده از این نتایج تولید سلول های بتا از سلول های بنیادی در شرایط آزمایشگاهی را تسهیل می کند.

    کلید واژگان: سلول های بنیادی مزانشیمی مغز استخوان, عصاره ی پانکراس, تمایز, سلول های انسولین ساز, سلول های بتای پانکراس
    Zahra Hatamnia, Fariba Esmaeili, Fariba Houshmand, Leila Dehdehi
    Background And Aims

    Diabetes type 1 is developed as a result of autoimmune destruction of pancreatic islet beta cells. Recent studies indicated that many types of stem cells could be considered ad potential sources of obtaining Insulin like producing cells (IPCS). In this study, the differentiation of bone marrow mesenchymal stem cells into insulin-producing cells was investigated using mouse pancreatic extract.

    Methods

    In this laboratory experimental study, bone marrow mesenchymal stem cells were used to differentiate into insulin-producing cells. The differentiated cells were assessed using dithizone-specific staining and anti-insulin-proinsulin and anti-insulin receptor beta antibodies. In addition, specific gene expression of pancreatic cells was assessed by RT-PCR method. Resaults: The differentiated cells derived from direct method of morphology were represented as pancreatic beta cells. Positive dithizone cells were observed as red-purple bundles. The results of RT-PCR represented specific gene expression of beta cells in the differentiated cells as direct. Immunofluorescence confirmed the presence of beta cells specific markers in these cells.

    Conclusion

    The results of this study indicated that bone marrow mesenchymal stem cells could differentiate into insulin-producing cells in presence of pancreatic extract. Therefore, application of these results facilitates in vitro production of beta cells from stem cells.

    Keywords: Bone marrow mesenchymal stem cells, Pancreatic extract, Differentiation, Insulin, producing cells, Pancreatic beta cells
نکته
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