26 research outputs found
The effect of Mesenchymal Stem Cells of Amniotic Membrane on the Proliferation and Differentiation of Umbilical Cord Blood CD34+ cells
Introduction: Ex vivo proliferation of hematopoietic stem cells (HSCs) of umbilical cord is widely used by combination of cytokine and stromal mesenchymal stem cells (MSCs) as feeder layer due to increase the cell doses, adequately. However, numerous studies have shown that ex vivo proliferation of these cells impairs their functions, including reduced self-renewal ability, apoptosis induction, and disordered cell cycle. MSCs have different sources such as amniotic membrane with a stable karyotype and high quality because of isolation from embryonic tissues, so that they are considered as a useful source for MSCs.Materials and Methods: In this study, isolated mesenchymal cells from the amniotic membrane were used as feeders for the HSCs proliferation. Four different cultures with various conditions were used; first one containing cytokines (stem cell factor, thrombopoietin, and FMS-related tyrosine kinase 3 ligand), second one with MSCs co-cultured with the aforementioned cytokines, third medium co-cultured with MSCs without cytokines, and finally the control medium was without co-culture condition and cytokines. Expression of mRNAs of HOXB4, GATA2, BCL2, and Survivin genes was also investigated.Results: The findings showed that the expression of mRNAs of these genes decreased in culture with cytokine, solely; however the expression of these genes was significantly higher in co-cultured system with cytokine rather than just with cytokine.Conclusion: : In general, the findings of this study indicate that the derived MSCs from amniotic membrane is a good source for the proliferation of umbilical cord blood CD34+ cells”. Because these cells increase the UCB-CD34+ quantity and their preservation properties.
Effects of maternal thyroid hormone deficiency on differentiation of mesenchymal stem cells in CSF-exposed neonatal Wistar rats
Impact of coadministration of apigenin and bone marrow stromal cells on damaged ovaries due to chemotherapy in rat: An experimental study
Background: Apigenin is a plant-derived flavonoid with antioxidative and antiapoptotic effects. Bone marrow stromal cells (BMSCs) are a type of mesenchymal stem cells (MSCs) that may recover damaged ovaries. It seems that apigenin may promote the differentiation of MSCs.
Objective: The aim of this study was to investigate the effect of coadministration of apigenin and BMSCs on the function, structure, and apoptosis of the damaged ovaries after creating a chemotherapy model with cyclophosphamide in rat.
Materials and Methods: For chemotherapy induction and ovary destruction, cyclophosphamide was injected intraperitoneally to 40 female Wistar rats (weighing 180–200 gr, 10 wk old) for 14 days. Then, the rats were randomly divided into four groups (n = 10/each): control, apigenin, BMSCs and coadministration of apigenin and BMSCs. Injection of apigenin was performed intraperitoneally and BMSC transplantation was performed locally in the ovaries. The level of anti-mullerian hormone serum by ELISA kit, the number of oocytes by superovulation, the number of ovarian follicles in different stages by H&E staining, and the expression of ovarian Bcl-2 and Bax proteins by western blot were assessed after four wk.
Results: The results of serum anti-mullerian hormone level, number of oocytes and follicles, and Bcl-2/Bax expression ratio showed that coadministration of apigenin and BMSCs significantly recovered the ovarian function, structure, and apoptosis compared to the control, BMSC, and apigenin groups (p < 0.001).
Conclusion: The results suggest that the effect of coadministration of apigenin and BMSCs is maybe more effective than the effect of their administrations individually on the recovery of damaged ovaries following the chemotherapy with cyclophosphamide in rats.
Key words: Apigenin, Bone marrow stromal cells, Chemotherapy, Ovary, Regeneration
Prenatal kisspeptin antagonist exposure prevents polycystic ovary syndrome development in prenatally-androgenized rats in adulthood: An experimental study
Background: Increased levels of kisspeptin are associated with hypothalamus-pituitary- ovary axis dysfunction. It may lead to the development of polycystic ovary syndrome (PCOS).
Objective: We aimed to investigate the effect of prenatal kisspeptin antagonist exposure on the development of PCOS in prenatally androgenized rats in adulthood.
Materials and Methods: In this experimental study, pregnant rats were injected with free testosterone (T, 5 mg/day) or T+P271 (kisspeptin antagonist) on the 20th day of the pregnancy period (n = 5 in each group), while rats in the control group received solvent. Female offspring were examined in terms of anogenital distance (AGD), anovaginal distance (AVD), vaginal opening, serum total testosterone (TT) levels, ovarian follicles, and the regularity of estrous cycles in adulthood. AGD and AVD were measured using a vernier caliper. TT levels were measured using the enzyme-linked immunosorbent assay method. Ovaries were fixed in 10% formalin, tissue processing was done by a standard protocol, and then ovaries embedded in paraffin. 5 μmthickness ovarian sections mounted on a glass slide, deparaffinized, and stained using Harris’s Hematoxylin and Eosin Y.
Results: AGD, AVD (p < 0.001), TT levels (p = 0.02), and the numbers of preantral and antral follicles (p < 0.001) in the ovaries were significantly decreased in prenatally TP271- exposed rats compared to prenatally T-exposed rats. The age of vaginal opening was early in T-P271-exposed rats compared to prenatally T-exposed rats (p < 0.001). The number of corpora lutea was significantly increased in T-P271-exposed rats (p < 0.001). No cystic follicles were observed in the ovaries of prenatally T-P271-exposed rats. Prenatally T-P271-exposed rats had regular estrous cycles compared to prenatally T-exposed rats.
Conclusion: Prenatal exposure to kisspeptin antagonist can prevent PCOS development in prenatally androgenized rats in adulthood.
Key words: Androgen, Kisspeptin antagonist, Polycystic ovary syndrome, Rat
Citral effect in male NMRI mice nonalcoholic steatosis model: assessing biochemical and histological parameters and PPARα gene expression
Citral is a small molecule present in various citrus species, with reported anti-hyperlipidemic and antiinflammation effects. Here, the effect of intraperitoneal (IP) administration of citral is evaluated in a mouse model of non-alcoholic steatosis. Male NMRI mice were divided into the following groups (n = 12): normal control group (NC) receiving a normal diet; high-fat emulsion group (HF) receiving high fat diet for four weeks; positive control group (C+) receiving HF diet for four weeks and then shifted to normal diet with IP-administered silymarin (80 mg/kg) for four weeks; sham group receiving HF diet for four weeks and then shifted to normal diet for four weeks; and EC1, EC2, and EC3 groups receiving HF diet for four weeks and then shifted to normal diet with IP-administered citral doses of 5, 10, and 20 mg/kg, respectively. HF diet resulted in steatohepatitis with impaired lipid profile, high glucose levels and insulin resistance, impaired liver enzymes, antioxidants, adiponectin and leptin levels, decreased PPARα level, and fibrosis in the liver tissue. Upon treatment with citral, improvement in condition was observed in a dose-dependent manner—both at histological level and in the serum of treated animals. and the PPARα level was also increased
Effect of Exosomes Derived from Bone Marrow Mesenchymal Stem Cells on Ovarian Granulosa Cells of Immature NMRI Mice
Objective: In recent years, in vitro maturation (IVM) has become the focus of fertility maintenance, and infertilitytreatment. The aim of this study is development of oocytes during folliculogenesis and oogenesis is greatly influencedby the presence of BMP-7, BMP-15, and GDF-9 genes, which are present in exosomes generated from bone marrowstem cells.Materials and Methods: In the experimental study, we investigated how exosomes obtained from bone marrow stemcells affected development and expansion of ovarian granulosa cells (GCs) in NMRI mice. In this in vitro experiment,bone marrow stem cells were isolated from mice’s bone marrow, and after identification, exosomes were recovered.Exosome doses of 100, 50, and 25 μg/ml were applied to GCs before using MTT assay to measure survival rates andquantitative reverse-transcription polymerase chain reaction (PCR) to measure expression of the BMP-7, BMP-15, andGDF-9 genes.Results: The results showed that the GCs treated with exosomes concentrations of 25, 50, and 100 μg/ml significantlyincreased bioavailability, growth and proliferation and it also increased expression level of BMP-7, BMP-15 and GDF-9genes compared to the controls.Conclusion: Findings of this study indicated that exosomes derived from bone marrow stem cells improved growthof GCs in NMRI mice and they were a good candidate for further clinical studies to improve quality of the assistedreproductive techniques
Protective Effect of Docetaxel Against Autophagy-Related Genes in Vitrification of Mouse Metaphase II Oocytes
Background: Autophagy is a conservative mechanism for cell survival as the main response of cells to stress conditions. The present study aimed to assess the effect of docetaxel on the survival, fertilization, and expression of autophagy-related genes in vitrified oocytes. Methods: The study was conducted in 2018 at the Stem Cells Technology Research Center, Shiraz University of Medical Sciences (Shiraz, Iran). Denuded oocytes were randomly selected and assigned to five groups, namely control (n=133), docetaxel (n=136), docetaxel+cryoprotectants (n=146), docetaxel+vitrification (n=138), and vitrification (n=145). The effect of vitrification on the expression of autophagy-related gene 5 (ATG5) and Beclin-1 was determined using a real-time polymerase chain reaction. Data were analyzed using SPSS software (version 26.0) and GraphPad Prism 9.Results: Survival and fertilization rates in each experimental group were significantly reduced compared to the control group (P=0.001). After in vitro fertilization of oocytes, the 2-cell formation rate was significantly reduced in the docetaxel+vitrification and vitrification groups compared to the control and docetaxel groups (P=0.001 and P=0.001, respectively). Pre-incubation of oocytes with docetaxel reduced gene expression levels of Beclin-1 and ATG5 in the docetaxel+cryoprotectants and docetaxel+vitrification groups (P=0.001 and P=0.019, respectively). The expression level of these genes was also reduced in the docetaxel group compared to the control group (P=0.001). Conclusion: Incubation of mouse metaphase II oocytes with docetaxel prior to vitrification reduced the expression of autophagy-related genes and increased survival and fertilization rates compared to untreated oocytes
Association of Cytochrome P450 2E1 and N-Acetyltransferase 2 Genotypes with Serum Isoniazid Level and Anti-Tuberculosis Drug-Induced Hepatotoxicity: A Cross-Sectional Study
Background: Anti-tuberculosis drug-induced hepatotoxicity can result from genetic polymorphism of the isoniazid (INH) metabolizing enzyme. This study aimed to determine the effect of genetic polymorphism of N-acetyltransferase 2 (NAT2) and cytochrome P450 2E1 (CYP2E1) genes on serum isoniazid level and drug-induced hepatotoxicity. Methods: A cross-sectional study was conducted on 120 patients (with and without hepatotoxicity) with pulmonary tuberculosis from June 2019 to April 2022 in Tehran (Iran). High-performance liquid chromatography was used to measure the serum concentration of INH and acetylisoniazid (AcINH). NAT2 and CYP2E1 genotypes were determined using polymerase chain reaction and restriction fragment length polymorphism methods. Data were analyzed using SPSS software (version 22.0) with independent two-sample t test, Chi square test, or Fisher’s exact test. P<0.05 was considered statistically significant.Results: A total of 40 patients showed hepatotoxicity. The risk of anti-tuberculosis drug-induced hepatotoxicity was significantly higher in patients who are slow acetylator (SA) phenotype than in rapid or intermediate acetylator (P<0.001). NAT2*4/*4 genotypes were not found in patients with hepatotoxicity. The frequency of NAT2*5 and NAT2*6 haplotypes and serum INH concentration was significantly higher in patients with hepatotoxicity than in those without (P=0.003, P<0.001, and P<0.001, respectively). NAT2*4 haplotype was correlated with protection against hepatotoxicity. A combination of SA and CYP2E1 C1/C1 genotype was significantly associated with hepatotoxicity (P<0.001). Conclusion: Hepatotoxicity in Iranian patients with tuberculosis was confirmed due to the presence of NAT2 SA polymorphism. Determining NAT2 and CYP2E1 genotypes and/or INH concentration can be a valuable tool to identify patients susceptible to hepatotoxicity
Comparison of viability of adipose-derived Mesenchymal stem cells on agarose and fibrin glue scaffolds
Background & aim: Utilizing tissue engineering techniques and designing similar structures of the damaged tissues require the use of tools such as scaffolds, cells, and bioactive molecules in vitro. Meanwhile, appropriate cell cultures with the ability to divide and differentiate on the natural scaffolds lacking features like immunogenicity and tumorgenesis is particularly important. Adipose tissue has attracted researchers’ attention due to its abundance of mesenchymal stem cells and its availability through a liposuction. The purpose of the present study was to investigate the reproducibility and viability of the adipose-derived stem cells on natural scaffolds of fibrin glue and agarose. Methods: In the present experimental study, the isolation and identification of the mesenchymal stem cells was performed on tissue obtained from liposuction. The tissues were extensively washed with PBS and were digested with collagenase I, then the mesenchymal stem cells were isolated. The cells were cultured in RPMI medium supplemented with antibiotic. Subsequently, the expression of cell surface markers including CD34, CD44, CD90, and CD105 were analyzed by flow cytometry to confirm the mesenchymal cells. After preparing fibrin glue and agarose scaffolds, the viability and proliferation of the adipose tissue-derived mesenchymal stem cells were examined at the period of 24, 48, and 72 hours by MTT and ELISA assays. The obtained results were analyzed by SPSS ver.19. Results: The results of adipose tissue-derived mesenchymal stem cells culture on the fibrin glue and agarose scaffolds indicated that cell viability on fibrin glue and agarose scaffold were 68.22% and 89.75% in 24 hrs, 64.04% and 66.97% in 48 hours, 222.87% and 1089.68% in 72 hours respectively. Significant proliferation and viability cells on a synthesized agarose scaffold were seen compared to the fibrin glue scaffold after 72 hrs. The viability of the cells significantly increased on the agarose scaffold. Conclusion: Due to stability and permeability of scaffolding agarose, it seems that scaffolding agarose created better adhesion of cells in the performance of cell proliferation process compared with fibrin glue scaffold
Lack of relatedness between Human cytomegalovirus in semen and male infertility
Background and Aim: Infertility refer to an inability to conceive (irrespective of causes) after having regular unprotected sex. Men are responsible for infertility in 50% of infertile spouses. Numerous factors contribute to male infertility including genital infections that may appear following microbial, fungal, and viral infections. The aim of the present study was detection of Human cytomegalovirus (HCMV) infection in the semen of infertile men and evaluation of its relation with sperm parameters such as motility, count and morphology.
Materials and Methods: In this study, 100 semen samples of fertile men (as a control group) and 100 semen samples of infertile men were collected from Infertility Center of Qom Jahad Daneshgahi, Iran. The semen samples were analyzed according to World Health Organization’s standard methods (WHO). After DNA extraction, cytomegalovirus was detected using Nested polymerase chain reaction technique. The results were analyzed using SPSS (version 19) software (Significance level of P<0.05). Statistical assessments were conducted using descriptive statistics including the Chi-square test and t test. The difference was considered significant at the P<0.05 level.
Results: The analyzed sperm parameters of 100 infertile samples showed that 56%, 70%, and 82% of the samples had problems, respectively in terms of count, motility, and morphology. However, 60% of the infertile samples had problems in all three parameters. Human cytomegalovirus DNA was detected in six and four samples of infertile and fertile men respectively.
Conclusions: According to statistical analysis, no significant correlation was obtained between the cytomegalovirus infection and male infertility. Although the achieved results may vary with changes in population size