Cell Journal (Yakhteh)
Volume:15 Issue: 4, Winter 2014

Stem cell therapy is a powerful technique for the treatment of a number of diseases. Stem cells are derived from different tissue sources, the most important of which are the bone marrow (BM), umbilical cord (UC) blood and liver. Human UC mesenchymal stem cells (hUC-MSCs) are multipotent, non-hematopoietic stem cells that have the ability to self-renew and differentiate into other cells and tissues such as osteoblasts, adipocytes and chondroblasts. In a number of reports, human and mouse models of disease have hUC-MSCs treatments. In this article, we review studies that pertain to the use of hUC-MSCs as treatment for diseases.

This study examines the effects of hydrostatic pressure on in vitro maturation (IVM) of oocytes derived from in vitro grown follicles. In this experimental study، preantral follicles were isolated from 12-day-old female NMRI mice. Each follicle was cultured individually in Alpha Minimal Essential Medium (α-MEM) under mineral oil for 12 days. Then، follicles were induced for IVM and divided into two groups، control and experiment. In the experiment group follicles were subjected to 20 mmHg pressure for 30 minutes and cultured for 24-48 hours. We assessed for viability and IVM of the oocytes. The percentage of apoptosis in cumulus cells was determined by the TUNEL assay. A comparison between groups was made using the student’s t test. The percentage of metaphase II oocytes (MII) increased in hydrostatic pressure-treated follicles compared to controls (p<0. 05). Cumulus cell viability reduced in hydrostatic pressure-treated follicles compared to controls (p<0. 05). Exposure of follicles to pressure increased apoptosis in cumulus cells compared to controls (p<0. 05). Hydrostatic pressure، by inducing apoptosis in cumulus cells، participates in the cumulus oocyte coupled relationship with oocyte maturation.

Nutrients and antioxidants in the medium of immature oocyte have a profound effect on maturation، fertilization and development of resulting embryos. In this study the effects of melatonin as an antioxidant agent on maturation، glutathione level and expression of high mobility group box-1 (HMGB1) gene were evaluated in immature oocytes of mice stained with brilliant cresyl blue (BCB). In this experimental study، immature oocytes were harvested from ovaries of Naval Medical Research Institute (NMRI) mice. Oocytes were stained with 26 μM BCB for 90 minutes and transferred to in vitro maturation medium containing varying doses of melatonin (10-12، 10-9، 10-6، 10-3 M) and without melatonin، for 22-24 hours. Maturation was monitored using an inverted microscope. Glutathione was assessed by monochlorobimane (MCB) staining and HMGB1 expression in mature oocyte was analyzed using real-time polymerase chain reaction (PCR). Melatonin in the concentration of 10-6 M had the most effect on maturation and HMGB1 expression of BCB+ oocytes (p<0. 05). Meanwhile melatonin had no effects on glutathione levels. Additionally in immature BCB- oocytes، compared to the control group، melatonin did not affect cytoplasm maturation (p>0. 05). In vitro treatment with melatonin increases the maturation and HMGB1 expression in BCB+ immature oocytes and has no significant effect on glutathione levels.

Oral mucosal epithelia of smokers and waterpipe users are more susceptible to malignant alterations. The aim of this study was morphometric evaluation of the effects of using waterpipe on normal oral mucosa. In a cross sectional study، cytologic smear samples from the following three different areas: buccal mucosa، lateral surface of the tongue، and floor of the mouth (right) were taken from 40 smokers، 40 waterpipe users، and 40 normal individuals. They were then stained using Papanicolaou staining technique. Quantitative cytologic alterations such as nuclear and cytoplasmic size، nuclear-cytoplasmic (N/C) ratio، Feret ratio (FR)، percent of karriorhexis، vacuolization of cytoplasm، two or multilobed nuclei، inflammation، and candida were evaluated. Quantitative evaluation was performed using MoticPlus 2 software، and 50 cells in each slide were studied. Practitioners were matched with age and sex in three groups. An increase in nuclear size، the N/C ratio، and F. R، while a decrease in cytoplasm size were observed in lateral surface of the tongue، buccal mucosa and floor of the mouth of smokers، waterpipe users and normal individuals، respectively (p≤0. 001). No statistically significant differences were observed in percent of karriorhexis، vacuolization of cytoplasm، and two or multilobed nuclei in oral mucosa of smokers، waterpipe users (p=0. 8)، and normal individuals (p=0. 9) in buccal mucosa،tongue، and mouth floor areas. However، the percentage of inflammation and candida in smokers (p<0. 001) and waterpipe users (p=0. 002) were higher than normal. individuals Smoking and using waterpipe are effective in creating some quantitative cytometric alterations in oral mucosa; however، smoking shows greater effect in the cytometric alterations than using waterpipe. Role of cytology in screening and detection of oral mucosa malignancies in smokers and waterpipe users needs further studies.

The effects of exposure to electromagnetic fields (EMF) on reproduction systems have been widely debated. In this study, we aimed to investigate whether low frequency EMF could ameliorate the in vitro fertilization success rate in Naval medical research institute (NMRI) Mice. In this randomized comparative animal study, ten NMRI mice were randomly divided into 2 equal groups (control and experimental). 10 IU of human chorionic gonadotropin (hCG) was injected intraperitoneally to both groups in order to stimulate ovulating, and ovums were then aspirated and kept in KSOM (modified version of sequential simplex optimization medium with a higher K+ concentration) culture medium. Metaphase II ovums were separated, and sperms obtained by «swim out» method were added to metaphase II ovums in the culture medium. The experimental group was exposed to 1. 3 millitesla pulsed electromagnetic field at 4 kilohertz frequency for 5 hours. To assess the efficacy, we considered the identification of two-pronuclear zygote (2PN) under microscope as fertilizing criterion. Total number of collected ovums in the control and experimental groups was 191 and 173, respectively, from which 58 (30. 05%) and 52 (30. 36%) ovums were collected from metaphase II, respectively. In vitro fertilization (IVF) success rate was 77% in extremely low frequency- pulsed electromagnetic field (ELF-PEMF) for exposed group (experimental), whereas the rate was 68% for control group. Despite increased percentile of IVF success rate in exposed group, there was no statistically significant difference between 2 groups, but this hypothesis has still been stated as a question. Further studies with larger sample sizes and different EMF designs are suggested.

Nuclear transfer-embryonic stem cells (NT-ESCs) are genetically identical to the donor’s cells; provide a renewable source of tissue for replacement, and therefore, decrease the risk of immune rejection. Trichostatin A (TSA) as a histone deacetylase inhibitor (HDACi) plays an important role in the reorganization of the genome and epigenetic changes. In this study, we examined whether TSA treatment after somatic cell nuclear transfer (SCNT) can improve the developmental rate of embryos and establishment rate of NT-ESCs line, as well as whether TSA treatment can improve histone modification in NT-ESCs lines. In this experimental study, mature oocytes were recovered from BDF1 [C57BL/6×DBA/2) F 1 mice] mice and enucleated by micromanipulator. Cumulus cells were injected into enucleated oocytes as donor. Reconstructed embryos were activated in the presence or absence of TSA and cultured for 5 days. Blastocysts were transferred on inactive mouse embryonic fibroblasts (MEF), so ESCs lines were established. ESCs markers were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Histone modifications were analyzed by enzyme linked immunosorbent assay (ELISA). Result of this study showed that TSA treatment after SCNT can improve developmental rate of embryos (21.12 ± 3.56 vs. 8.08 ± 7.92), as well as establishment rate of NT-ESCs line (25 vs. 12.5). We established 6 NT-ESCs in two experimental groups, and three embryonic stem cells (ESCs) lines as control group. TSA treatment has no effect in H3K4 acetylation and H3K9 tri-methylation in ESCs. TSA plays a key role in the developmental rate of embryos, establishment rate of ESC lines after SCNT, and regulation of histone modification in NT-ESCs, in a manner similar to that of ESCs established from normal blastocysts.

In this study, artificial neural network (ANN) analysis of virotherapy in preclinical breast cancer was investigated. In this research article, a multilayer feed-forward neural network trained with an error back-propagation algorithm was incorporated in order to develop a predictive model. The input parameters of the model were virus dose, week and tamoxifen citrate, while tumor weight was included in the output parameter. Two different training algorithms, namely quick propagation (QP) and Levenberg-Marquardt (LM), were used to train ANN. The results showed that the LM algorithm, with 3-9-1 arrangement is more efficient compared to QP. Using LM algorithm, the coefficient of determination (R2) between the actual and predicted values was determined as 0.897118 for all data. It can be concluded that this ANN model may provide good ability to predict the biometry information of tumor in preclinical breast cancer virotherapy. The results showed that the LM algorithm employed by Neural Power software gave the better performance compared with the QP and virus dose, and it is more important factor compared to tamoxifen and time (week).

Research related to induce pluripotent stem (iPS) cell generation has increased rapidly in recent years. Six transcription factors, namely OCT4, SOX2, C-MYC, KLF4, NANOG, and LIN28 have been widely used for iPS cell generation. As there is a lack of data on intra- and inter-networking among these six different transcription factors, the objective of this study is to analyze the intra- and inter-networks between them using bioinformatics.In this computational biology study, we used AminoNet, MATLAB to examine networking between the six different transcription factors. The directed network was constructed using MATLAB programming and the distance between nodes was estimated using a phylogram. The protein-protein interactions between the nuclear reprogramming factors was performed using the software STRING. The relationship between C-MYC and NANOG was depicted using a phylogenetic tree and the sequence analysis showed OCT4, C-MYC, NANOG, and SOX2 together share a common evolutionary origin. This study has shown an innovative rapid method for the analysis of intra and inter-networking among nuclear reprogramming factors. Data presented may aid researchers to understand the complex regulatory networks involving iPS cell generation.

The aim of the study was to compare the effects of two different concentrations of cryoprotectants by cryotopvitrification on survival, developmental capacity and Heat shock protein 72 (Hsp72) expression of two-cell mouse embryos. In this experimental study, transcript analysis of Hsp72 gene was performed on non-vitrified and vitrified 2-cell mouse embryos via a nested quantitative polymerase chain reaction (nqPCR) subsequent to normalization with Hprt1 as the reference gene. The different cryoprotectant combinations were 15% (vit1:7.5% of each ethylene glycol (EG) and dimethyl sulfoxide (DMSO), 30% (vit2:15% EG + 15% DMSO) and control group with no cryoprotectants. Vitrified and fresh 2-cell embryos were cultured to obtain cleavage and blastocyst formation rates. The results were analyzed via one-way analysis of variance and the mean values were compared with least significant difference (LSD) (p< 0.05). The relative expression of Hsp72 in vit2 (30% v/v) was significantly higher than vit1 (15% v/v). Survival rates were the same for both vitrification treatments and significantly lower than the control group. Cleavage and blastocyst rates in vit1 were significantly higher than vit2 while those in two vitrified groups were significantly lower than the control group. Our developmental data demonstrated that vit1 treatment (7.5% EG and 7.5% DMSO) was more efficient than vit2 (15% EG and 15% DMSO) in mouse embryos. The cryotopvitrification with two concentrations of cryoprotectants caused the relative changes of Hsp72 transcript level, but the stability of the gene in vit1 was significantly higher than vit2 and closer to the fresh 2-cell embryos.

Methoxsalen is a natural photoactive compound which is found in many seed plants. A number of epidermal proliferative disorders can be treated by methoxsalen along with long wave ultraviolet A (UVA). In an experimental study, we aimed to demonstrate the effect of methoxsalen, UVA and their combination on oogenesis Balb/C mice. There were two experimental groups and a control group. The experimental groups were composed of i. a short term group with treatment duration of 15 days and ii. a long term group with treatment duration of 5 weeks. Both the long term and short term experimental groups were further subdivided into a UVA group, a methoxsalen group and a methoxsalen plus UVA group. After treatment, mature females in prosterus phase of ovarian cycle were scarified with ether, while their ovaries were removed and prepared for histological studies. Both macro and microscopic studies showed significant anomalies (p<0.05) among experimental group ovaries as compared to control group. The obtained results showed a significant decrease in the following factors: number and diameter of corpus lutei, Graafian follicles, diameter of granulosa cell layer and oocytes, number of primordial،and primary and growing follicles, while we observed an increase in number of atretic follicle. Furthermore, our findings confirmed an increase in theca diameter only through UVA treatment. Methoxsalen also reduced circulating estrogen levels in blood serum, significantly. Other cases of teratogenecity, such as follicles with three oocytes and disorganization in corpus luteum cells were observed. The result suggests that UVA, methoxsalen and their combination cause health problems and cell injuries.

To evaluate the mRNA expression ratio of Bcl-2/Bax both in normal and tumoral bladder tissues of patients with transitional cell carcinoma (TCC) of bladder and investigate potential correlation between this expression ratio and clinical outcome. In this experimental study, we used real time-PCR to investigate the expression of Bcl-2 and Bax both in normal and tumoral bladder tissues. The Bcl-2/Bax expression ratio was determined in tumoral bladder tissues of patients with transitional cell carcinoma of the bladder (n=40) and correlation between expression ratios and the emergence of early relapses in a follow-up of 14-30 months was examined. Relapse-free time in 14/31 patients (45.16%) with Bcl-2/Bax>1 was shorter than 9 months (range of 2-9 months) with 5.7 months average median while 17/31 patients (54.84%) with Bcl-2/Bax<1 are currently relapse-free (14-30 months). Bcl-2 and Bax expression levels were not solely correlated with clinical outcome and progression of carcinogenesis. The mRNA expression ratio of Bcl-2/Bax in tumoral bladder tissues may serve as a significant prognostic indicator in predicting the clinical outcome in low grade non-invasive bladder cancer.

Recently, the use of stem cells has expanded into numerous areas including cell therapy. In this study, we investigated the differentiation capacity of human Wharton’s jelly stem cells (hWJSCs) into lens fiber cells. Morphological changes and expressions of four crystallin genes (αA, αB, βB1 and βB3) were studied. The bovine vitreous body has been shown to induce expression of crystallin genes in hWJSCs. By using the vitreous as a lens fiber cell inducer, we showed that αB-, βB1- and βB3-crystallin genes expressed in hWJSCs.

One of the most significant steps in gene expression studies is transferring genes into cell cultures. Despite there are different methods for gene delivery such as viral and non-viral producers, some cationic lipid reagents have recently developed to transfect into mammalian cell lines. The main aim of this study was optimizing and improving lipocomplex based transient transfection procedures into HeLa cell line which is being used widely as a typical cell in biological studies.This study was an experimental research. In this work, pCMV: β-Gal DNA plasmid was used as a reporter DNA for determining the rate of gene transfection into HeLa cells. To accomplish the highest gene delivery into HeLa cells, optimizing experiments were carried out in different volumes of FuGENE-HD, LipofectamineTM2000 and X-tremeGENE. Also, we investigated tranasfection efficiency in presence of various cell densities of HeLa cells. Then, transfection efficiency and cell toxicity were measured by beta gal staining and trypan blue methods, respectively.Using FuGENE-HD in volume of 4μl along with 105 HeLa cells, transfection efficiency was higher (43.66 ± 1.52%) in comparison with the cationic lipids lipofectamineTM2000 and X-tremeGENE. In addition, the rate of cell toxicity in presence of FuGENE-HD was less than 5%. In summary, the cationic lipid FuGENE-HD indicates a suitable potential to transfer DNA into HeLa cells and it can be an efficient reagent for gene delivery for HeLa cells in vitro. Moreover, it is worth designing and optimizing gene transfer experiments for other cell lines with FuGENE-HD due to its low toxicity and high efficiency.