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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: VETERINARY RESEARCH FORUM
- نوع مقاله: Journal Article
- کلمات کلیدی: Testosterone,Extracellular matrix,Bovine testicular cells
- چکیده:
- چکیده انگلیسی: Testosterone is believed to play a significant role in spermatogenesis, but its contribution to the process of spermatogenesis is not completely understood. Given that extracellular matrix (ECM) facilitates differentiation of spermatogonial stem cells (SSCs) during culture, the present study was conducted to elucidate whether testosterone contribute to the permissive effect of ECM on SSCs differentiation. In experiment 1, testosterone production was measured in testicular cells cultured for 12 days on ECM or plastic (control). In experiment 2, testosterone production was assessed in testicular cells cultured on ECM or plastic (control) and exposed to different concentrations of hCG. In experiment 3, the gene expression of factors involved in testosterone production was analyzed. Testosterone concentration was lower in ECM than in the control group in experiment 1 (p < 0.05). In experiment 2, testosterone concentration was increased in response to hCG in both groups but cells cultured on ECM were more responsive to hCG than those cultured on plastic (p < 0.05). In the experiment 3, qRT-PCR revealed the inhibitory effect of ECM on the gene expression of steroidogenic acute regulatory protein (StAR) (p < 0.05). Nevertheless, the expression of LH receptor was greater in ECM-exposed than in unexposed cells (p < 0.05). In conclusion, the present study showed that inhibiting the expression of StAR, ECM could lower testosterone production by Leydig cells during in vitro culture. In addition, the results indicated that ECM could augment the responsiveness of Leydig cells to hCG through stimulating the expression of LH receptor.
- انتشار مقاله: 24-11-1395
- نویسندگان: Vahid Akbarinejad,Parviz Tajik,Mansoureh Movahedin,Reza Youssefi
- مشاهده
- جایگاه : پژوهشی
- مجله: VETERINARY RESEARCH FORUM
- نوع مقاله: Journal Article
- کلمات کلیدی: bovine,Differentiation,Spermatogonial stem cells,Fibroblast growth factor receptors
- چکیده:
- چکیده انگلیسی: The receptors 1 and 2 of fibroblast growth factor (FGFR1 and FGFR2, respectively) have been observed in all types of testicular cells. Culture on extracellular matrix (ECM) has been observed to lead to initiation of differentiation in spermatogonial stem cells (SSCs). The present study was carried out to investigate whether FGFR1 and FGFR2 play a role in SSCs differentiation. Following isolation, bovine testicular cells were cultured on ECM-coated or uncoated (control) plates for 12 days. The gene expression of THY1, cKIT, FGFR1 and FGFR2 was evaluated using quantitative real-time polymerase chain reaction (PCR). Results related to the gene expression of markers of with undifferentiated (THY1) and differentiated (cKIT) spermatogonia implicated stimulation of self-renewal and differentiation in cells cultured on ECM-coated and uncoated plates, respectively (p < 0.05). Concomitantly, the expression of FGFR2 increased during culture in the ECM group (p < 0.05), whereas it did not change in the control group (p > 0.05). As a result, the gene expression of FGFR2 was greater in the ECM than control group (p < 0.05). Nevertheless, FGFR1 expression did not change during culture in the control and ECM groups (p > 0.05). In conclusion, the present study revealed the potential role of FGFR2 in differentiation of SSCs during culture on ECM.
- انتشار مقاله: 09-03-1395
- نویسندگان: Vahid Akbarinejad,Parviz Tajik,Mansoureh Movahedin,Reza Youssefi
- مشاهده
- جایگاه : پژوهشی
- مجله: VETERINARY RESEARCH FORUM
- نوع مقاله: Journal Article
- کلمات کلیدی: bovine,Co-culture,FSH,Sertoli,SSCs
- چکیده:
- چکیده انگلیسی: The complex process of spermatogenesis is regulated by various factors. Studies on spermatogonial stem cells (SCCs) have provided very important tool to improve herd genetic and different field. 0.2 to 0.3 percent of total cells of seminiferous tubules is consist of spermatogonial stem cells. To investigate and biomanipulation of these cells, proliferation and viability rate of cells should be increased in vitro, at first. Follicle stimulating hormone (FSH) has been suggested to play a determinant role in the survival of germ cells in addition to increasing spermatogonial proliferation. In this study, the in vitro effects of FSH on spermatogonial cell colony formation were investigated. Sertoli and spermatogonial cells were isolated from 3-5 months old calves. The identity of the Sertoli cells and spermatogonial stem cells were confirmed through immunocytochemistry and colony morphology, respectively. Co-cultured Sertoli and spermatogonial cells were treated with FSH in different dose of 10, 20 and 40 IU mL-1 FSH, before colony assay. Results indicated that, FSH increased in vitro colonization of spermatogonial cells in comparison with control group. In conclusion, using FSH provided proper bovine spermatogonial stem cell culture medium for in vitro study of these cells.
- انتشار مقاله: 24-12-1391
- نویسندگان: Reza Narenji Sani,Parviz Tajik,Mohammad Hassan Yousefi,Mansoureh Movahedin,Babak Qasemi-Panahi,Shiva Shafiei,Mahmood Ahmadi Hamedani
- مشاهده
- جایگاه : پژوهشی
- مجله: Journal of Applied Biotechnology Reports
- نوع مقاله: Journal Article
- کلمات کلیدی: Ginseng,Infertility,Free radicals,Freezing and Thawing,Active Mitochondria of Sperm
- چکیده:
- چکیده انگلیسی: Introduction: Today, freezing method is one of the most common approaches in the treatment of infertility. This is while oxidative stress is a major destructing factor during sperm freezing and thawing. However, antioxidant compounds can play a key role in sperm freezing techniques. Materials and Methods: In this study, 5 groups of sperms were evaluated with and without Ginseng extract and then some parameters were evaluated such as mobility, activity of mitochondrial, amount of reactive oxygen species (ROS) and DNA fragmentation. Results: Findings revealed that the mobility and mitochondrial activity in sperms significantly increased (P≤0.05) in frozen and thawed sperms, which had been treated with 1 mg/mL Ginseng extract compared to the freezing and thawing sperms without Ginseng extract. In addition, the results showed that Ginseng treatment significantly decreased the amount of ROS and DNA fragmentation compared to freezing and thawing in frozen and thawed treatment without Ginseng (P ≤ 0.05). By preventing the increase of oxidative stress levels, Ginseng prevented the reduction of mobility and mitochondria activity of the sperms after freezing and thawing. It also reduced sperm DNA defeat and reduced the production of free radicals. Conclusions: The results of the present study support the hypothesis that Ginseng has positive effects on the sperm quality in cryopreservation process.
- انتشار مقاله: 09-02-1397
- نویسندگان: Arash Hasirbaf-Momtaz,Mansoureh Movahedin,Zohreh Mazaheri
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Veterinary Medicine
- نوع مقاله: Journal Article
- کلمات کلیدی: Spermatogonial stem cell,CSF1,bovine spermatogonia,SSC coculture,SSC niche
- چکیده: زمینه مطالعه: سلولهای بنیادی اسپرماتوگونی (SSCs) گروه کوچکی از سلولهای اسپرماتوگونی نوع A هستند که می توانند سلولهایی کاملاً مشابه خود ایجاد نموده (خودسازی) و همچنین تمایز یابند و در پایان روند اسپرماتوژنز، اسپرم را تولید نمایند. فرآیند خودسازی SSCs به طور کامل شناخته نشده است. برای فراهم شدن امکان مطالعه خصوصیات و عملکرد این سلولها که منجر به خودسازی و تمایز آنها می گردد، دسترسی به تعداد کافی از آنها ضروری است. هدف: در این مطالعه اثر دوزهای مختلف CSF1 روی کلونیزاسیون SSCs در همکشتی با سلولهای سرتولی بررسی شد. CSF1 از عواملی است که پیشنهاد شده ممکن است در SSC niche (جایگاه قرارگیری SSCs در لوله های سمینیفروس) که در آن خودسازی به تمایز برتری دارد، نقش داشته باشد. روش کار: تعداد و مساحت کلونی های اسپرماتوگونی ایجاد شده در حضور دوزهای متفاوت CSF1 (0، 10، 50 و 100 ng/ml) طی روزهای 4، 7 و 11 کشت ارزیابی گردید. توسط رنگ آمیزی ایمیونوسیتوفلورسنت علیه مارکرهای OCT4 و vimentin ماهیت SSCs و سلولهای سرتولی تاٌیید شد. نتایج: تعداد کلونی ها از روز 4 تا روز 11 در تمامی گروهها مستقل از حضور و یا دوز CSF1 به شکل معنی داری افزایش نشان داد. مشاهده گردید که تعداد کل کلونی ها و مجموع مساحت آنها در گروههای SCF1 10 و 50 ng/ml نسبت به گروههای CSF1 100 و 0 (کنترل) بالاتر است اما این تقاوت تنها در مورد مقایسه مجموع مساحت کلونی ها بین گروه CSF1 10 ng/ml و کنترل و تنها در روز 4 معنی دار بود (p
- چکیده انگلیسی: BACKGROUND: Spermatogonial stem cells (SSCs) are infrequent
self-renewing cells among the type A spermatogonia
within the seminiferous tubules and are the basis of spermatogenesis
in mammalian testis. An adequate number of SSCs is a
primary requirement for the study of their behavior, regulation, and
further biomanipulation. OBJECTIVES: In this paper, we studied
the development of the primary co-cultures of type A spermatogonia
and prepubertal bovine sertoli cells in the presence of Colony
Stimulating Factor 1 (CSF1), a potential contributor in the SSC
niche. METHODS: The effect of different concentrations of CSF1
(0, 10, 50 and 100 ng/mL) on the colonization activity of spermatogonial
cells was assessed 4, 7 and 11 days after the beginning of the
culture by counting the total number of colonies and measuring their
area in each group of the present experiment. Immunofluorescent
staining against OCT4 and vimentin led to the confirmation of the
nature of both the SSCs and sertoli cells. RESULTS: Results showed
that the total number of colonies from day 4 to 11 increased
significantly in all groups, independent of CSF1 concentration. In
addition, the total number and total area of colonies were higher (not
significant) in 10 and 50 ng/mL CSF1 treatments than the control
and 100 ng/mL CSF1 groups in all the three evaluations during the
experiment. However, this difference was only significant (p<0.05)
between the total area of colonies in the control and 10 ng/mLCSF1
groups at day 4 of co-culture. CONCLUSIONS:It was concluded that
CSF1 can be a suitable growth factor for improving SSCs colonization
in vitro, particularly during the first days of culture where
accompanying sertoli cells still have not proliferated sufficiently to
support the propagating spermatogonial cells.- انتشار مقاله: 15-08-1391
- نویسندگان: Shiva Shafiei,Parviz Tajik,Hamid Ghasemzadeh-nava,Mansoureh Movahedin,Massoud Talebkhan Garoussi,Babak Qasemi-Panahi,Peyman Rahimi Feyli
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Stem cells,Lithium chloride,Neuron-like cells,Induction,Transdifferentiation
- چکیده:
- چکیده انگلیسی: Objective(s): Adipose-derived stem cells (ADSCs), with suitable and easy access, are multipotential cells that have the ability for differentiation into other mesodermal and transdifferentiate into neural phenotype cells. In this study, Lithium chloride (LiCl) was used for in vitro transdifferentiation of rat ADSCs into neuron-like cells (NLCs).
Materials and Methods: ADSCs were isolated from the rats’ perinephric region using Dulbecco΄s Modified Eagle΄s Medium (DMEM) with Fetal Bovine Serum (FBS), cultured for 3 passages, characterized by flowcytometry and differentiation into adipogenic and osteogenic phenotypes. The ADSCs were exposed to 0.1, 0.5, 1, 1.5, 2, 5, and 10 millimolar (mM) LiCl without serum for 24 hr. The optimum dose of LiCl was selected according the maximum viability of cells. The expression of neurofilament light chain (NfL), neurofilament high chain (NfH), and nestin was evaluated by immunocytochemistry. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the amount of synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes’ expression in ADSCs and NLCs.
Results: The optimum dose of LiCl was 1 mM in 24 hr. The transdifferentiated ADSCs showed cytoplasmic extension with synapse-like formation. Synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes were significantly expressed more in NLCs than in ADSCs.
Conclusion: LiCl can induce ADSCs into neural phenotype cells with higher expression of neural and neuronal genes.- انتشار مقاله: 12-04-1398
- نویسندگان: Samaneh Farrokhfar,Taki Tiraihi,Mansoureh Movahedin,Hossein Azizi
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Stem cells,Lithium chloride,Neuron-like cells,Induction,Transdifferentiation
- چکیده:
- چکیده انگلیسی: Objective(s): Adipose-derived stem cells (ADSCs), with suitable and easy access, are multipotential cells that have the ability for differentiation into other mesodermal and transdifferentiate into neural phenotype cells. In this study, Lithium chloride (LiCl) was used for in vitro transdifferentiation of rat ADSCs into neuron-like cells (NLCs).
Materials and Methods: ADSCs were isolated from the rats’ perinephric region using Dulbecco΄s Modified Eagle΄s Medium (DMEM) with Fetal Bovine Serum (FBS), cultured for 3 passages, characterized by flowcytometry and differentiation into adipogenic and osteogenic phenotypes. The ADSCs were exposed to 0.1, 0.5, 1, 1.5, 2, 5, and 10 millimolar (mM) LiCl without serum for 24 hr. The optimum dose of LiCl was selected according the maximum viability of cells. The expression of neurofilament light chain (NfL), neurofilament high chain (NfH), and nestin was evaluated by immunocytochemistry. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the amount of synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes’ expression in ADSCs and NLCs.
Results: The optimum dose of LiCl was 1 mM in 24 hr. The transdifferentiated ADSCs showed cytoplasmic extension with synapse-like formation. Synaptophysin, neurogenin-1, neuroD1, NfL, NfH, and nestin genes were significantly expressed more in NLCs than in ADSCs.
Conclusion: LiCl can induce ADSCs into neural phenotype cells with higher expression of neural and neuronal genes.- انتشار مقاله: 12-04-1398
- نویسندگان: Samaneh Farrokhfar,Taki Tiraihi,Mansoureh Movahedin,Hossein Azizi
- مشاهده