الگوی بیان چرخه‌های سیگنالی LIF و 2- FGF در سلول‌های بنیادی رویانی حاصل از لقاح آزمایشگاهی، همتاسازی و خودگشنی در گاومیش

نوع مقاله: مقاله کامل

نویسندگان

گروه علوم دام، طیور و آبزیان، پژوهشکده کشاورزی، سازمان پژوهش‌های علمی و صنعتی ایران، تهران، ایران

چکیده

سلول های بنیادی رویانی که از لایه زاینده داخلی رویان در مرحله بلاستوسیست تولید می شوند، توانایی تمایز به  انواع سلول های لایه زاینده رویان شامل برون پوست ، میان پوست و درون پوست را دارند. اطلاعات کمی در ارتباط با فاکتورهایی که بر تمایز یا حفظ سلول در حالت تمایز نیافته اثر می گذارند وجود دارد. در این مطالعه الگوی بیان چرخه های فاکتور مهارکننده لوکمیا (LIF) و فاکتور رشد فیبروبلاستی (2-FGF)، به منظور درک بهتر ارتباط چرخه های سیگنالی در خودنوسازی سلول های بنیادی رویانی گاومیش مورد بررسی قرار گرفت. سلول های بنیادی رویانی از رویان های حاصل از لقاح آزمایشگاهی (IVF)، همتاسازی  (IVC) و خودگشنی (Parthenogenesis) بدست آمدند و از آلکالین فسفاتاز و رنگ آمیزی ایمیونو فلورسانس به منظور شناسایی سلول های بنیادی استفاده شد. به منظور کشت سلول های بنیادی از لایه تغدیه کننده استفاده شد و محیط کشت حاوی Ko-DMEM،KSR ، LIF، 2-FGF، ال- گلوتامین، اسیدهای آمینه غیر ضروری و جنتامایسین بود. بیان ژن های حد واسط چرخه های LIF و 2-FGF با استفاده از RT-PCR بدست آمد. نتایج حاصل از این مطالعه نشان داد، بیان2-FGF در سلول های بنیادی رویانی گاومیش بیشتر از LIF بود. 2-LIF ، FGF، گیرنده های آن ها و ترکیبات حد واسط این چرخه ها در سلول های بنیادی رویانی حاصل از هر سه منشاء تقریبا در حد یکسانی بیان شدند.

کلیدواژه‌ها


عنوان مقاله [English]

Transcriptional profiling of LIF and FGF-2 signaling pathways in buffalo embryonic stem cells derived from in vitro fertilized, parthenogenetic and handmade cloned embryos

نویسندگان [English]

  • M. Zandi
  • M.R. Sanjabi
Iranian Research Organization for Science and Technology (IROST).
چکیده [English]

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of blastocyst and differentiate into all three embryonic germ layers: ectoderm, endoderm, and mesoderm. There is less information available about the factors that are affecting buffalo ES cells in culture. In this study, expression profiles of the Leukemia Inhibitory Factor (LIF) and basic Fibroblast Growth Factor (FGF-2) signaling pathways were investigated to better understand the relationships of the signaling pathways for self-renewal in buffalo ES cells. Buffalo ES cells were derived from in vitro fertilized (iESC), parthenogenetic (pESC) and handmade cloned (cESC) embryos. Alkaline phosphatase and immune-fluorescence staining were used to characterize buffalo ES cells. Feeder layer was used for ESCs culture, and culture medium consisting of Knockout- Dulbecco’s Modified Eagle’s Medium (Ko-DMEM) supplemented with Knockout Serum Replacement (KSR), leukemia inhibitory factor (LIF), basic fibroblast growth factor-2 (FGF-2), L-glutamine, nonessential amino acids and gentamicin. Gene expression was analysed by RT-PCR for signaling pathways. Results showed that, the expression of FGF-2 was higher than LIF in buffalo ESCs. LIF, FGF, their receptors and intermediate signaling pathways was expressed at almost same level in three sources of buffalo ESCs.

کلیدواژه‌ها [English]

  • Embryonic Stem Cells
  • FGF
  • LIF
  • Buffalo
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