تولید سلول های بنیادی پرتوان القایی از سلول های خون محیطی با استفاده از ریزمولکول ها

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه زیست شناسی سلولی و مولکولی، دانشکده علوم زیستی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران

2 گروه زیست شناسی سلول و مولکولی، دانشکده علوم و فناوری های زیستی، دانشگاه شهید بهشتی، تهران، ایران

3 آزمایشگاه طب بازساختی و نوآوری های زیست پزشکی، انستیتو پاستور ایران، تهران، ایران

4 گروه بانک سلولی، انستیتو پاستور ایران، تهران، ایران

10.22034/AEJ.2021.297827.2595

چکیده

تولید سلول­ های بنیادی پرتوان القایی با استفاده از روش ­های ایمن و غیرویروسی نقش به­ سزایی در مهندسی بافت و طب بازساختی دارد. در این تحقیق سعی شد تا سلول ­های بنیادی پرتوان القایی را در حضور انواعی از ریزمولکول­ های شیمیایی از سلول­ های خون محیطی تولید نمود. سلول ­های تک هسته ­ای خونی با استفاده از روش فایکول جداسازی و تکثیر آن­ ها توسط لیپوپلی ساکارید القاء گردید. بازبرنامه ­ریزی لنفوسیت­ های استخراج شده با استفاده از ترکیبی از ریزمولکول­ ها مانند آزاسیتیدین، سدیم بوتیرات، کنپائولون و رپساکس که عمدتاً مدولاتورهای اپی­ ژنتیکی موثر بر ژن ­های دخیل در پرتوانی (Oct4،Sox2 ،Klf4 ، C-Myc) صورت پذیرفت. در ادامه سلول­ های بنیادی پرتوان القایی حاصله بر روی لایه تغذیه کننده فیبروبلاست جنین موش انتقال داده شده و مورد شناسایی قرار گرفتند. لیپوپلی ساکارید به ­طور چشمگیری منجر به افزایش تعداد سلول­ های خونی تک هسته­ ای بعد از 5 ساعت گردید. سلول­ های خونی پس از 14 روز تیمار با ترکیبات شیمیایی و ریزمولکول ­های مورد نظر به­ طور موفقیت ­آمیزی به سلول­ های بنیادی پرتوان القایی تبدیل شدند و سپس جهت تکثیر و حفظ خاصیت پرتوانی بر روی محیط حمایتی فیبروبلاست جنین موش قرار گرفتند. استفاده از ترکیبات شیمیایی و ریزمولکول­ ها روش ایمن و ارزان برای تولید موثر سلول­ های بنیادی پرتوان القایی از سلول ­های خونی می­ باشد و به ­کارگیری فیبروبلاست جنین موش به­ عنوان لایه تغذیه­ کننده فاکتورهای رشد مورد نیاز جهت حفظ حالت پرتوانی را تولید می­ کند.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Generation of induced pluripotent stem cells from peripheral blood cells using small molecules

نویسندگان [English]

  • Javad Kazemi 1
  • Hosein Shahsavarani 2 3
  • Parviz Pakzad 1
  • Mohammadali Shokrgozar 3 4
1 Department of Cellular and Molecular Biology, Faculty of Biological Sciences, North Tehran Branch, Islamic Azad University, Tehran, Iran
2 Department of Cell and Molecular Sciences, Faculty of Life Science and Biotechnology, Shahid Beheshti University, Tehran ،Iran|Laboratory of Regenerative Medicine and Biomedical Innovation, Pasteur Institute of Iran, Tehran, Iran
3 Department of Cell and Molecular Sciences, Faculty of Life Science and Biotechnology, Shahid Beheshti University, Tehran ،Iran|Laboratory of Regenerative Medicine and Biomedical Innovation, Pasteur Institute of Iran, Tehran, Iran
4 Laboratory of Regenerative Medicine and Biomedical Innovation, Pasteur Institute of Iran, Tehran, Iran|Department of National Cell Bank, Pasteur Institute of Iran, Tehran, Iran
چکیده [English]

Generation of induced pluripotent stem cells (iPSCs) using safe and non-viral methods play pivotal role in tissue engineering and regenerative medicine. In current study, we aimed to generate iPSCs from peripheral blood mononuclear cells (PBMC) in the presence of different small molecule and chemical compounds. PBMCs were isolated using Ficoll gradient method and their proliferation was induced by lipopolysaccharide (LPS). Reprogramming of extracted PBMCs was mediated using a combination of small molecule such as 5-Azacitidine, Kenpaullone, sodium butyrate and RepSox, which are generally epigenetic modulator of genes (OCT4, Sox2, Klf4, c-Myc) involved on pluripotent cell. Then, resulted pluripotent stem cells were cultured and identified on feeder MEFs. LPS effectively caused an increase in the number of PMBCs after 5 hours. Blood cells were successfully converted to iPSCs after treatment with different chemical compounds and small molecules for 14 days and then transferred on supportive mouse embryonic fibroblasts. The use of small molecules is a safe and cost-effective method for generating iPSCs from PBMCs. Mouse embryonic fibroblasts serve as a feeder layer for production of growth factors and preserving pluripotency of stem cells.

کلیدواژه‌ها [English]

  • Induced pluripotent stem cells
  • Mouse embryonic fibroblasts
  • Chemical small molecule compounds
  • Blood cells

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فصلنامه محیط زیست جانوری
دوره 14، شماره 2
تیر 1401
صفحه 267-276

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