ABSTRACT
Human adipose stem cells (hASCs) were introduced as appropriate candidate due to advantages like ease of isolation, in vitro expansion and lack of immune response. Deprenyl (Dep) was used to induce bone marrow stem cells into neuron-like cells. We investigated the Dep effect on neurotrophin genes expression in hASCs and their differentiation into neuron-like cells. The cells were isolated from small pieces of abdominal adipose tissue and subjected to flow cytometry to confirm purification. The osteogenic and adipogenic differentiation were identified. The proliferation rate and neurotrophin genes expression of treated cells were evaluated by MTT, TH immunostaining and RT-PCR. hASCs had positive response to CD44, CD73, CD90, CD105 markers and negative response to CD34 and CD45 markers and differentiated into adipocytes and osteocytes. Exposure to 10–7 M of Dep for 24 hours caused a significant increase of viable cells and BDNF, NTF-3 genes expression as compared to cultured cells in serum free medium and had no effect on the expression of NGF and GDNF genes. Based on our results, Dep is able to induce BDNF, NTF-3 and NTF-4 genes expression and neroun-like morphology in hASCs.
ABSTRACT
Human adipose stem cells (hASCs) were introduced as appropriate candidate due to advantages like ease of isolation, in vitro expansion and lack of immune response. Deprenyl (Dep) was used to induce bone marrow stem cells into neuron-like cells. We investigated the Dep effect on neurotrophin genes expression in hASCs and their differentiation into neuron-like cells. The cells were isolated from small pieces of abdominal adipose tissue and subjected to flow cytometry to confirm purification. The osteogenic and adipogenic differentiation were identified. The proliferation rate and neurotrophin genes expression of treated cells were evaluated by MTT, TH immunostaining and RT-PCR. hASCs had positive response to CD44, CD73, CD90, CD105 markers and negative response to CD34 and CD45 markers and differentiated into adipocytes and osteocytes. Exposure to 10–7 M of Dep for 24 hours caused a significant increase of viable cells and BDNF, NTF-3 genes expression as compared to cultured cells in serum free medium and had no effect on the expression of NGF and GDNF genes. Based on our results, Dep is able to induce BDNF, NTF-3 and NTF-4 genes expression and neroun-like morphology in hASCs.
ABSTRACT
Background: One of the most major obstacles of ovarian tissue vitrification is suboptimal developmental competence of follicles. Matrix metalloproteinases 2 [MMP-2] and 9 [MMP-9] and their tissue inhibitors TIMP-1 and TIMP-2 are involved in the remodeling of the extracellular matrix in the ovaries
Objective: This study aimed to evaluate the expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 genes in the preantral follicles derived from vitrified mouse ovaries
Materials and Methods: In this experimental study, the gene expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the isolated preantral follicles derived from fresh and vitrified ovaries of 14-16 days old female mice through real time qRT-PCR was evaluated. Developmental parameters, including survival rate, growth, antrum formation and metaphase II oocytes were also analyzed
Results: The developmental parameters of fresh preantral follicles were significantly higher than vitrified preantral follicles. The TIMP-1 and MMP-9 expression levels showed no differences between fresh and vitrified preantral follicles [p=0.22, p=0.11 respectively]. By contrast, TIMP-2 expression significantly decreased [p=0.00] and MMP-2 expression increased significantly [p=0.00] in vitrified preantral follicles compared with to fresh ones
Conclusion: Changes in expression of MMP-2 and TIMP-2 after ovarian tissues vitrification is partially correlated with decrease in follicle development
ABSTRACT
Background: The uterus is a dynamic tissue responding to hormonal changes during reproductive cycles. As such, uterine stem cells have been studied in recent years. Transcription factors oct4 and sox2 are critical for effective maintenance of pluripotent cell identity
Objective: The present research evaluated the mRNA expression of oct4 and sox2 in the uterine tissues of ovariectomized mice treated with steroid hormones
Materials and Methods: In this experimental study, adult virgin female mice were ovariectomized and treated with estradiol 17 [E2], progesterone [P4], and a combination of E2 and P4 [E2 and P4] for 5 days. Uterine tissues were removed, and immunofluorescent [IF] staining and quantitative real-time PCR of oct4 and sox2 markers were performed
Results: IF showed oct4 and sox2 expression in the uterine endometrium and myometrium among all groups. The mRNA expression of oct4 [p=0.022] and sox2 [p=0.042] in the E2-treated group significantly were decreased compared to that in the control group. By contrast, the mRNA expression of oct4 and sox2 in the P4 [p=0.641 and 0.489 respectively] and E2 and P4-treated groups [p=0.267 and 0.264 respectively] did not show significant differences compared to the control group
Conclusion: The results indicate ovarian steroid hormones change the expression of oct4 and sox2 in the mice uterine tissues, which suggest the involvement of steroid hormonal regulation in uterine stem cells
ABSTRACT
Since the uterine is a sensitive tissue to steroid hormones, the aim of the present study was to investigate the effects of 17 beta-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
ABSTRACT
The kiwi fruit is known to have dramatic antibacterial, debridement, wound contracture, and angiogenic effects. We propose that kiwifruit is an ideal candidate to enhance the process of wound healing. The present study assessed the effects of wound kiwifruit dressing on cutaneous wound healing in rat. In this experimental study, 30 male Wistar rats were randomly divided into 2 groups of control and kiwifruit group. A full-thickness dermal incision [35 mm length] was made on the right side of the paravertebral region. In the control group, one day after wound induction wounds was dressed with vaseline sterile gauze after normal saline irrigation. In the second group, the wounds were dressed with kiwifruit. Wound healing was evaluated by measuring surface area, percentage of healing, duration of healing, and wound tensile strength. Obtained results showed that the duration of wound healing in kiwi group in comparison with the control was significantly decreased. The amount of wound healing in percent was also significantly different between control and kiwi groups at days 3, 6 [p<0.001], 9 [p<0.05], 12 and 14 [p<0.01]. Comparisons of wound length between control and kiwi group per day showed that kiwi group had significantly lower wound length on day 9, 12, 14 and 16 [p<0.01, 0.001, 0.01 and 0.01, respectively] in comparisons to control group. Also, the wound tensile strength in kiwi group also was significantly greater than the control animals [p<0.01]. We concluded that our study provides some evidence to support the use of kiwi to accelerate wound healing
ABSTRACT
Cryopreservation of ovarian tissues and pre-antral follicles is a promising prospect for preservation of women fertility. The aim of this study was to evaluate the in vitro developmental competence of mouse vitrified pre-antral follicles in comparison to isolated pre-antral follicles derived from vitrified ovaries in the presence of alpha lipoic acid [ALA]. Pre-antral follicles derived from fresh, vitrified-warmed ovarian tissues and vitrified-warmed pre-antral follicles were cultured individually with or without ALA, followed by adding hCG to induce ovulation. The follicle growth, oocyte maturation, and embryo development were assessed. The diameter and development of follicles, oocyte maturation and embryo development rates were significantly higher in ALA supplemented groups compared to the respective ALA-free conditions groups. Aforementioned parameters were significantly higher in vitrified-warmed follicles in comparison to follicles derived from vitrified-warmed ovaries. These findings support a superior performance of pre-antral follicles when vitrified rather than when isolated from vitrified ovaries with regard to increasing the rates of developmental parameters. Moreover, ALA improves the in vitro maturation of pre-antral follicles in vitrified and non-vitrified samples.
ABSTRACT
Cryopreservation of pre-antral follicles is a hopeful technique to preserve female fertility. The aim of the present study was to evaluate reactive oxygen species [ROS] and total antioxidant capacity [TAC] levels of mouse vitrified pre-antral follicles in the presence of alpha lipoic acid [ALA]. Isolated pre-antral follicles [140-150 micro m in diameter] were divided into vitrified-warmed and fresh groups. Each group was subjected to in vitro maturation with or without ALA for 12 days, followed by adding human chronic gonadotropin to induce ovulation. In vitro fertilization was performed to evaluate their developmental competence. In parallel, the amount of ROS and TAC were assessed after 0, 24, 48, 72, and 96 h of culture by 2',7'-dichlorofluorescin assay and ferric reducing/antioxidant power assay, respectively. The respective rates of survival, antrum formation, and metaphase II oocytes were significantly higher in ALA-supplemented groups compared to the groups not treated with ALA. TAC and ROS levels were significantly decreased and increased, respectively during the culture period up to 96 h in the absence of ALA in both vitrified and non-vitrified samples. However, with pretreatment of ALA, TAC levels were increased significantly and remained constant up to 96 h in vitrified-warmed pre-antral follicles, while ROS levels completely returned to the level of starting point after 96 h of culture in the presence of ALA. Pretreatment of ALA positively influences development of pre-antral follicles in vitrified and nonvitrified samples through increasing follicular TAC level and decreasing ROS levels
ABSTRACT
There is longstanding experimental and clinical evidence that supports the idea that replacement of dopaminergic [DAergic] neurons can ameliorate functional disabilities of Parkinson's disease [PD]. The purpose of the present study is to examine the efficacy of transplantation of rat bone marrow stromal cell [BMSCs]-derived tyrosine hydroxylase-positive [TH[+]] cells induced by deprenyl into 6-hydroxydopamine [6-OHDA]-lesioned rat models, using behavioral tests and immunohistochemical evaluations. In this experimental study, undifferentiated BrdU-labeled BM-SCs were incubated in serum-free medium that contained 10[-8] M deprenyl for 24 hours. Afterwards, BMSCs were cultured for 48 hours in alpha-minimal essential medium [alpha-MEM] supplemented with 10% FBS, then differentiated into Th[+] neurons. We randomly divided 24 hemiparkinsonian rats as follows: group 1 [control] received only medium, while groups 2 and 3 were injected with 2x10[5] BMSCs and deprenyl-treated cells in 4 microl medium. Injections were made into the injured strata of the rats. Rotational behavior in response to apomorphine was tested before transplantation and at 2, 4, and 6 weeks post-graft. Animals were then sacrificed, and the brains were extracted for immunohistochemical and electron microscopic studies. Apomorphine-induced rotation analysis indicated that animals with grafted cells in groups 2 and 3 exhibited significantly less rotational behavior than those in the control group at 2, 4, and 6 weeks after transplantation. Immunohistochemical analysis demonstrated that BrdU-labeled cells expressed specific neuronal markers, such as NF 200 and TH, at the implantation site. The presence of TH[+] cells in conjunction with the reduction in rotation might show the capacity of grafted cells to release dopamine. Ultrastructural analysis revealed the presence of immature neurons and astrocyte-like cells at the graft site. Th[+] neurons induced by deprenyl can be considered as a cell source for PD autograft therapy?
Subject(s)
Male , Animals, Laboratory , Selegiline , Tyrosine 3-Monooxygenase , Oxidopamine , Rats , Models, Animal , Parkinson Disease , Immunohistochemistry , BehaviorABSTRACT
In spite of extensive efforts to improve in vitro oocyte maturation, the obtained results are not very efficient. This study was conducted to assess impacts of cAMP elevating agents and alpha lipoic acid [ALA] on in vitro oocyte maturation and fertilization. Mouse germinal vesicle [GV] oocytes were categorized into cumulus denuded oocytes [DOs; n=896] and cumulus oocyte complexes [COCs; n=1077] groups. GV oocytes were matured in vitro with or without ALA; [I] without the meiotic inhibitors; [II] supplemented with cilostamide; [III] supplemented with forskolin and [IV] supplemented with Forskolin plus cilostamide. The obtained metaphase II [MII] oocytes were subjected to in vitro fertilization. Independent-samples t-test and ANOVA were used for data analysis. A p-value less than 0.05 was considered to be statistically significant. The COCs maturation, fertilization and two cell embryo rates were higher than those of DOs in the control group, while no significant difference was observed between relevant COCs and DOs when they were cultured with cilostamide meiotic inhibitors in two step manner. Combined treatment of cilostamide and forskolin significantly elevated the developmental rates in both COCs and DOs as compared to other groups. The developmental rates of COCs and DOs in the presence of ALA were similar to their respective groups without ALA. cAMP elevating agents were more effective on DOs than COCs with regard to rates of maturation and fertilization. However, ALA did not affect the developmental rates of both COCs and DOs in in vitro maturation in one or two step manner
Subject(s)
Female , Animals, Laboratory , In Vitro Oocyte Maturation Techniques , Cyclic AMP , Mice , Cumulus Cells , OocytesABSTRACT
It has been reported that rat bone marrow stromal cells [BMSCs] can be spontaneously differentiated into neural-like cells without any supplemental growth factors and/or chemical treatment after long-term culture. This study aims to determine whether, growth factors secreted by MSCs could induce self-differentiation into neural-like cells in a long-term culture. This study consisted of two groups: i. rat BMSCs [passage 5] were cultured in alfa- minimal essential medium [alpha-MEM] and 10% fetal bovine serum [FBS] without the addition of inducer and exchanging medium for three weeks, as the experimental group and ii.rat BMSCs [passage 5] as the control group. Each group was analysed by reverse transcriptase polymerase chain reaction [RT-PCR] to evaluate the expressions of neurotrophic factors and neural marker genes. Statistical analyses were carried out using one-way analysis of variance [ANOVA] and Tukey's multiple comparison with SPSS software [version 16]. P< 0.05 was considered statistically significant. The experimental group [fifth passage of BMSCs] obtained from adult rats spontaneously differentiated into neural precursor cells after long-term culture. Cultured cells expressed tyrosine hydroxylase [TH], Nurr1 and nestin genes. Furthermore, some growing cells 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]. In this study, we reported that rMSCs in long-term culture underwent spontaneous transformation to neural precursors without the supplement of growth factors and specific chemicals. Cells expressed neural markers such as: TH, Nurr1, and nestin genes
Subject(s)
Animals, Laboratory , Cell Culture Techniques , Nerve Tissue Proteins , Bone Marrow Cells , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , ImmunohistochemistryABSTRACT
This research study is an attempt to examine whether the administration of ethanol after memory reactivation would modulate subsequent expression of memory in rats. Additionally, we examined whether this administration alters the density of Cornu Ammonis [CA]1 and CA3 pyramidal and dentate gyrus [DG] granule cells. In this experimental study, adult male Wistar rats [200-300 g] were trained in a fear conditioning system using two 1 second, 0.6 mA shocks with an interval of 180 seconds. Twenty four hours later rats were returned to the chamber for 120 seconds. Immediately after reactivation they were injected with ethanol [0.5, 1, 1.5 mg/ kg] or saline. 1, 7 and 14 days after reactivation, rats were returned to the context for 5 minutes. Seconds of freezing [absence of all movement except respiration] were scored. In the second experiment [described in the previous paragraph], after test 1, animals were anesthetized with sodium pentobarbital and perfused transcardially with phosphate buffer [10 minutes] and 4% paraformaldehyde [15 minutes]. The brains were postfixed in phosphate-buffered 4% paraformaldehyde [24 hours] and 30% sucrose. 10-?m sections were stained with cresyl violet. Data were analyzed by 1-and 2-way ANOVA for repeated measurements by means of SPSS 16.0. Tukey's post hoc test was performed to determine the source of detected significant differences. P <0 .05 were considered significant. Data are presented as mean +/- SEM. Findings from the first experiment indicated that ethanol at a dose of 1.5 mg/kg significantly impaired recall of memory only in the first test. The density of CA1 and CA3 pyramidal and DG granule cells in the ethanol group was decreased [p< 0.01] compared with control group respectively 43.7%, 35.8%, and 37.8. The data demonstrate that ethanol exposure impairs post retrieval processes. Moreover, ethanol decreases the density of CA1, CA3 and DG cells. Presumably it would be a correlation between our behavioral and histological results
Subject(s)
Animals, Laboratory , Ethanol , Rats, WistarABSTRACT
This study is an attempt to examine the transdifferentiation of bone marrow stromal cells [BMSCs] into tyrosine hydroxylase immunoreactive cells in parkinsonian rats associated with angiogenesis. In this study, Sprague-Dawley rats received unilateral stereotaxic injections of 6-hydroxydopamine[6-OHDA] into the left corpus striatum and then were divided into two groups. One group, the negative control, received only medium while the other group was treated with BMSCs. BMSCs were harvested from femur bones, labeled with bromodeoxyuridine [BrdU] and then transplanted into parkinsonian rats, where a behavioral study and immunohistochemistry were used to evaluate the treatment, The results showed statistically significant improvement in rotational behavior. Anti-BrdU antibody showed engraftment of the transplanted cells at the transplantation site. Additionally, double immunolabeling confirmed that these cells were positive for neurofilament-200 and tyrosine hydroxylase [TH]. It may be concluded that BMSCs transplants could engraft and differentiate into TH immunoreactive cells which may cause recovery from motor deficits. Also, BMSCs may contribute to angiogenesis at the transplantation site