بررسی اثر هم‌کشتی سلولهای بنیادی بر بافت تخمدان موش سوری
محورهای موضوعی : زیست شناسیعلی محمد عینی 1 , احمدعلی محمدپور 2 , عباس پرهام 3
1 - دانشجوی دکتری، گروه بافت‌شناسی، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران
2 - استاد، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران
3 - دانشیار، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه فردوسی مشهد، مشهد، ایران
کلید واژه: بافت تخمدان, سلولهای بنیادی مزانشیمی, کشت بافت, موش سوری, کشت سلول,
چکیده مقاله :
هدف: با استفاده از تکنیک‌های کشت سلول و کشت بافت، بسیاری از محققان برای روشن کردن عوامل هورمونی لازم برای تمایز ساختاری و فعالیت‌های عملکردی بافت تخمدان تلاش می‌‌کنند. لذا، مطالعه حاضر به بررسی کشت بافت تخمدان بر روی سلولهای بنیادی مزانشیمی جدا شده از بافت چربی پرداخته است.مواد و روش‌ها: در این مطالعه سلولهای بنیادی مزانشیمی از بافت چربی محوطه بطنی موش سوری جدا گردید. پس از پنج روز و تشکیل تک لایه‌‌ای از سلولهای بنیادی مزانشیمی، تخمدان‌‌های گرفته شده به مدت هفت روز بر روی سلولهای کشت شده مزانشیمی قرار گرفت. داده‌های به دست آمده با استفاده از نرم‌افزار Spss تجزیه و تحلیل شد.یافته‌ها: نتایج بدست آمده در این مطالعه نشان داد که تعداد فولیکولهای بالغ گروه تیمار نسبت به گروه کنترل به ترتیب (9/1±9/38، 2/2±1/61) بوده است که در مقایسه با گروه کنترل افزایش معنی‌داری داشته است (P<0/05). این نتیجه نشان‌‌دهنده موثر بودن کشت بافت در هم‌کشتی با سلولهای بنیادی مزانشیمی می‌باشد. همچنین در بررسی داده‌های مربوط به آزمایش RT-PCR مقدار ژن BAX کاهش چشمگیری در گروه مورد مطالعه نسبت به گروه کنترل داشته است (P<0.05). با توجه به نتایج به دست آمده از تعداد و کیفیت فولیکولهای بدست آمده به نظر می‌‌رسد این روش در تولید فولیکولهای بالغ و متعاقب آن اووسیت‌‌های با کیفیت و تولید جنین کارآمد و دارای اهمیت فراوانی باشد.
Objective: Many researchers are trying to clarify necessary hormonal factors for structural distinguishability and functional activities of ovarian tissue by using cell culture and tissue culture techniques. Hence, in this study, we examined ovarian tissue culture on mesenchymal stem cells isolated from cultured adipose tissue.Materials and methods: In this study, mesenchymal stem cells became isolated from adipose tissue of the mouse. After five days and monolayer formation of Mesenchymal stem cells, obtained ovarian was placed on the cultured Mesenchymal stem cells’ monolayer for 7 days.Results: The results of this study show that numbers of mature follicles for treat group compared to the control group were 61/1±2/2 and 38/1±1/9, respectively which had a significant increase compared to control group (P<0/05). This shows the efficiency of tissue culture in co-culture with Mesenchymal stem cells. The real-time PCR studies also confirmed the decrease in the BAX genes in treatment Groups to compare the control group (P<0/05). According to the results yielded regarding the number and quality of obtained follicles, it seems that this method is efficient and of high importance in producing mature follicles, and subsequently high quality oocytes and embryo.
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Eppig, JJ & O'Brien, MJ. Development in vitro of mouse oocytes from primordial follicles. Biology of Reproduction.1996; 54(1): 197-207.
Kern S, Eichler H, Stoeve J, Klüter H & Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006; 24: 1294-301.
Babu, N & Nair NB. Follicular atresia in Amblypharyngodon Chakaiensis. Z. mikrosk.
- anat. Forsch. 1983; 97: 499-504.
Martinovitch P. The development in vitro of the mammalian gonad. Ovary and ovogenesis. Proceedings of the Royal Society of London. Series B. Biological Sciences. 1938; 125 (839): 232-249.
Hennet, ML & Combelles C. The antral follicle: a microenvironment for oocyte differentiation. The International journal of developmental biology. 2012; 56(3): 819-831.
Blandau RJ, Warrick E & RE. Rumery: In vitro cultivation of fetal mouse ovaries. Fert Steril.1965; 16: 705-715.
Klawitter M & et al. Curcuma DMSO extracts and curcumin exhibit an anti-inflammatory and anti-catabolic effect on human intervertebral disc cells, possibly by influencing TLR2 expression and JNK activity. Journal of Inflammation. 2012; 9: 29.
Terada N, Kuroda H, Namiki M, Kitamura Y & Matsumoto K. Augmentation of aromatase activity by FSH in ovaries of fetal and neonatal mice in organ culture. J Steroid Biochem.1984; 20: 741-745.
Neal P & Baker TG. Response of mouse ovaries in vivo and in organ culture to pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. I Examination of critical time intervals. J Reprod Fertil. 1973; 33: 399-410.
Neal P & Baker TG. Response of mouse ovaries in vivo and in organ culture to pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. II: Effect of different doses of hormones. J Reprod Fertil. 1974; 37: 399-404.
Funkenstein B, Nimrod A & Lindner HR. The development of steroidogenic capability and responsiveness to gonadotrophins in cultured neonatal rat ovaries. Endocrinology. 1980; 106: 98-106.
Henrotiny AL & et al. Mobasher, Biological actions of curcumin on articular chondrocytes. Osteoarthritis and Cartilage. 2010; 18, 141-149.
Raphael L & et al. Mesenchymal cell interaction with ovarian cancer cells induces a background dependent pro-metastatic transcriptomic profile. Journal of Translational Medicine. 2014; 12: 59.
Rasmusson I, Ringdén O, Sundberg B, Le Blanc K. Mesenchymal stem cells inhibit lymphocyte proliferation by mitogens and alloantigens by different mechanisms. Exp Cell Res. 2005; 305: 33-41.
Toghraie FS & et al. Treatment of osteoarthritis with infrapatellar fat pad derived mesenchymal stem cells in Rabbit. Knee. 2011; 18: 71-5.
Voge J & et al. Effect of insulin-like growth factors (IGF), FSH, and leptin on
IGF-binding-protein mRNA expression in bovine granulosa and theca cells: quantitative detection by real-time PCR. Peptides. 2004; 25(12): 2195-2203.
Nestler JE. Insulin regulation of human ovarian androgens. Human reproduction. 1997; 12 (suppl 1): 53-62.
Jin SY, & et al. A novel two-step strategy for in vitro culture of early-stage ovarian follicles in the mouse. Fertility and sterility. 2010; 93(8): 2633-2639.
Silva J, & et al. Control of oestradiol secretion and of cytochrome P450 aromatase messenger ribonucleic acid accumulation by FSH involves different intracellular pathways in oestrogenic bovine granulosa cells in vitro. Reproduction. 2006; 132(6):
909-917.
Acevedo N & Ding J. Insulin signaling in mouse oocytes. Biology of Reproduction. 2007; 77(5): 872-879.
_||_Lanza R & Atala A. Essential of Stem Cell Biology.Third Edition, Elsevier Academic Press, 2014.
Eppig, JJ & O'Brien, MJ. Development in vitro of mouse oocytes from primordial follicles. Biology of Reproduction.1996; 54(1): 197-207.
Kern S, Eichler H, Stoeve J, Klüter H & Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006; 24: 1294-301.
Babu, N & Nair NB. Follicular atresia in Amblypharyngodon Chakaiensis. Z. mikrosk.
- anat. Forsch. 1983; 97: 499-504.
Martinovitch P. The development in vitro of the mammalian gonad. Ovary and ovogenesis. Proceedings of the Royal Society of London. Series B. Biological Sciences. 1938; 125 (839): 232-249.
Hennet, ML & Combelles C. The antral follicle: a microenvironment for oocyte differentiation. The International journal of developmental biology. 2012; 56(3): 819-831.
Blandau RJ, Warrick E & RE. Rumery: In vitro cultivation of fetal mouse ovaries. Fert Steril.1965; 16: 705-715.
Klawitter M & et al. Curcuma DMSO extracts and curcumin exhibit an anti-inflammatory and anti-catabolic effect on human intervertebral disc cells, possibly by influencing TLR2 expression and JNK activity. Journal of Inflammation. 2012; 9: 29.
Terada N, Kuroda H, Namiki M, Kitamura Y & Matsumoto K. Augmentation of aromatase activity by FSH in ovaries of fetal and neonatal mice in organ culture. J Steroid Biochem.1984; 20: 741-745.
Neal P & Baker TG. Response of mouse ovaries in vivo and in organ culture to pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. I Examination of critical time intervals. J Reprod Fertil. 1973; 33: 399-410.
Neal P & Baker TG. Response of mouse ovaries in vivo and in organ culture to pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. II: Effect of different doses of hormones. J Reprod Fertil. 1974; 37: 399-404.
Funkenstein B, Nimrod A & Lindner HR. The development of steroidogenic capability and responsiveness to gonadotrophins in cultured neonatal rat ovaries. Endocrinology. 1980; 106: 98-106.
Henrotiny AL & et al. Mobasher, Biological actions of curcumin on articular chondrocytes. Osteoarthritis and Cartilage. 2010; 18, 141-149.
Raphael L & et al. Mesenchymal cell interaction with ovarian cancer cells induces a background dependent pro-metastatic transcriptomic profile. Journal of Translational Medicine. 2014; 12: 59.
Rasmusson I, Ringdén O, Sundberg B, Le Blanc K. Mesenchymal stem cells inhibit lymphocyte proliferation by mitogens and alloantigens by different mechanisms. Exp Cell Res. 2005; 305: 33-41.
Toghraie FS & et al. Treatment of osteoarthritis with infrapatellar fat pad derived mesenchymal stem cells in Rabbit. Knee. 2011; 18: 71-5.
Voge J & et al. Effect of insulin-like growth factors (IGF), FSH, and leptin on
IGF-binding-protein mRNA expression in bovine granulosa and theca cells: quantitative detection by real-time PCR. Peptides. 2004; 25(12): 2195-2203.
Nestler JE. Insulin regulation of human ovarian androgens. Human reproduction. 1997; 12 (suppl 1): 53-62.
Jin SY, & et al. A novel two-step strategy for in vitro culture of early-stage ovarian follicles in the mouse. Fertility and sterility. 2010; 93(8): 2633-2639.
Silva J, & et al. Control of oestradiol secretion and of cytochrome P450 aromatase messenger ribonucleic acid accumulation by FSH involves different intracellular pathways in oestrogenic bovine granulosa cells in vitro. Reproduction. 2006; 132(6):
909-917.
Acevedo N & Ding J. Insulin signaling in mouse oocytes. Biology of Reproduction. 2007; 77(5): 872-879.