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ارزیابی تأثیر فاکتور رشد Activin A و کوچک مولکول IDE1 در تکثیر و خودنوزایی سلولهای زایای بدوی جوجه در شرایط کشت آزمایشگاهی | ||
| زیست شناسی کاربردی | ||
| مقاله 7، دوره 36، شماره 3 - شماره پیاپی 77، آذر 1402، صفحه 69-85 اصل مقاله (1.37 M) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22051/jab.2023.42190.1523 | ||
| نویسندگان | ||
| معصومه زارع1؛ سیدضیالدین میرحسینی* 2؛ سیده نفیسه حسنی3؛ شاهرخ قوتی4 | ||
| 1دانشجوی دکتری، گروه علوم دامی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران. | ||
| 2استاد، گروه علوم دامی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران. | ||
| 3دانشیار.پژوهشگاه رویان، پژوهشکده زیستشناسی و فناوری سلولهای بنیادی جهاد دانشگاهی، مرکز تحقیقات علوم سلولی، گروه سلولهای بنیادی و زیستشناسی تکوینی، تهران، ایران | ||
| 4استادیار ،گروه علوم دامی، دانشکده علوم کشاورزی، دانشگاه گیلان، رشت، ایران | ||
| چکیده | ||
| مقدمه: چالش اصلی سلولها سلولهای زایای بدوی (PGC)، عدم تکثیر و خودنوزایی آنها در محیط کشت است. یک روش برای القای پرتوانی سلولهای PGC، دستکاری مسیرهای پیامرسانی درونسلولی ازجمله TGF-β است که استفاده از فاکتورهای رشد و کوچک مولکولها یکی از روشهای رسیدن به این هدف است. مواد و روش: سلولهای PGC گناد جوجه با غلظت 50000 سلول به ازای هر خانه در پلیت 24 خانه کوت شده با ماتریژل کشت و انکوبه شدند. گروههای آزمایشی شامل چهار گروه: کنترل (محیط پایه برای کشت PGCs)، تیمار با کوچک مولکول IDE1 (100 nM; Stemgent, USA, 04-0026)، تیمار با فاکتور رشد A Activin (25 ng/ml; R&D Systems, 338-AC) و تیمار با SB431542 (10 µM; Cayman Chemical, 13031) با سه تکرار از هر گروه بودند. به منظور بررسی میزان تکثیر سلولی، شمارش سلولهای PGC در بازههای زمانی 7، 14 21 روز بعد از تیمار با هموسایتومتر انجام شد. فعالیت مسیر سیگنالینگ TGF/ẞ با بررسی بیان ژنهای SMAD2، SMAD3 وLFTTY1 با روش qRT-PCR ارزیابی شد. نتایج: تأثیر Activin A و IDE1 منجر به افزایش تکثیر سلولهای PGCs به بیش از 4 برابر در مقایسه با گروه کنترل گردید و در مقابل گروه SB431542 منجر به کاهش تکثیر سلولی شد. گروههای آزمایشی Activin A و IDE1 قادر به حفظ زندهمانی و کشت سلولها به مدتزمان 25 روز شدند، اما منجر به کشت طولانیمدت نگردید. همچنین نتایج بررسی فعالیت مسیر سیگنالینگ TGF/ẞ نشان داد که Activin A و IDE1 منجر به افزایش بیان ژنهای Smad2، Smad3 و LFTTY1 در مقایسه با گروههای کنترل و SB431542شدند. | ||
| کلیدواژهها | ||
| پرتوانی؛ خودنوزایی؛ سلولهای زیای بدوی؛ Activin A؛ کوچک مولکول IDE1 | ||
| عنوان مقاله [English] | ||
| Evaluation of the effect of growth factor Activin A and small molecule IDE1 on the proliferation and self-renewal of cells Primitive chick embryo in laboratory culture conditions | ||
| نویسندگان [English] | ||
| Masoumeh Zareh1؛ Seyed Ziaedin Mirhoseini2؛ Seyedeh Nafiseh Hasani3؛ Shahrokh Ghovvati4 | ||
| 1PhD Student, Department of Animal Science, Faculty of Agricultural Science, University of Guilan, Rasht, | ||
| 2Professor, Department of Animal Science, Faculty of Agricultural Science, University of Guilan, Rasht, Iran | ||
| 3Associate Professor. Royan Research Institute, Jihad University Institute of Stem Cell Biology and Technology, Cell Science Research Center, Department of Stem Cells and Developmental Biology, Tehran, Iran | ||
| 4Assistant Professo.Department of Animal Science, Faculty of Agricultural Science, University of Guilan, Rasht, Iran | ||
| چکیده [English] | ||
| Introduction: The main challenge of primordial germ cells (PGC) is their lack of proliferation and self-renewal in the culture medium. One method for inducing the pluripotency of PGC cells is to manipulate intracellular signaling pathways such as TGF-β, and the use of growth factors and small molecules is one of the ways to achieve this goal. Materials and methods: Chicken gonadal PGC cells were cultured and incubated with a concentration of 50,000 cells per well in a 24-well plate coated with Matrigel. The experimental groups included four groups: control (basic medium for PGCs culture), treatment with small molecule IDE1 (100 nM; Stemgent, USA, 04-0026), treatment with growth factor A Activin (25 ng/ml; R&D Systems, 338-AC) and treatment with SB431542 (10 µM; Cayman Chemical, 13031) with three replicates from each group. In order to check the amount of cell proliferation, PGC cells were counted in time intervals of 7, 14, 21 days after the treatment with a hemocytometer. The activity of TGF/ẞ signaling pathway was evaluated by examining the expression of SMAD2, SMAD3 and LFTTY1 genes by qRT-PCR method. Results: The effect of Activin A and IDE1 led to an increase in the proliferation of PGCs cells to more than 4 times compared to the control group, and in contrast to the SB431542 group, it led to a decrease in cell proliferation. | ||
| کلیدواژهها [English] | ||
| Pluripotency, self-renewal, primitive germ cells, Activin A, small molecule IDE1 | ||
| مراجع | ||
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