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بررسی اثر محیط کشت رویی سلولهای بنیادی مزانشیمی جداشده از کلیه موش نوزاد بر روی تمایز سلولهای بنیادی جنینی انسان به سلولهای پیش ساز کلیه | ||
| فصلنامه علمی زیست شناسی جانوری تجربی | ||
| دوره 12، شماره 1 - شماره پیاپی 45، شهریور 1402، صفحه 99-113 اصل مقاله (1.7 M) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.30473/eab.2023.68943.1923 | ||
| نویسندگان | ||
| فاطمه رازقی1؛ مرجان نوری2؛ فتاح ستوده نژادنعمت الهی1؛ احسان احسانی* 3؛ کاظم پریور1 | ||
| 1گروه زیستشناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران | ||
| 2گروه علوم صنایع غذایی، واحد رودهن، دانشگاه آزاد اسلامی، رودهن، ایران | ||
| 3گروه زیستشناسی، واحد رودهن، دانشگاه آزاد اسلامی، رودهن، ایران | ||
| چکیده | ||
| بیماریهای کلیه به عنوان یک مشکل مهم پزشکی، دارای گزینههای محدود برای درمان است، بکارگیری سلول های بنیادی به عنوان یک روش درمانی جایگزین روشهای قدیمی مطرح است. هدف مطالعه حاضر بررسی اثر محیط کشت حاصل از سلولهای بنیادی مزانشیمی (MSCs-CM) کلیه موش نوزاد در درصدهای 10، 30 و 50 بر تمایز سلولهای بنیادی جنینی به سمت سلولهای اپیتلیال کلیه است. سلولهای بنیادی مزانشیمی، از کلیه موش نوزاد جداسازی، پاساژ و تکثیر شد. تعیین هویت سلولها با استفاده از فلوسایتومتری و بررسی بیان مارکرهای سطحی CD105، CD29، CD90، انجام شد. در پاساژ سوم سلولهای استخراج شده، محیط کشت رویی آن جمعآوری و hESC در محیط کشت کامل این سلولها کشت و تکثیر شدند و تمایز سلولهای hES به سلولهای پیش ساز مورد بررسی قرار گرفت. بررسی بیان ژنهای PAX2، ZO1 وCK18 با استفاده از RT-PCR و بیان مارکرهای سطحی CD133، CD24 و CD44 با استفاده از فلوسایتومتری بررسی شد. نتایج فلوسایتومتری ماهیت مزانشیمی بودن سلولها را تایید کرد. نتایج تمایز hESCs نشان داد که بیان ژنهای PAX2، ZO1 و CK18 در گروههای حاوی محیط کشت رویی به صورت معنیدار (p<0.05) افزایش یافتهاست. نتایج فلوسایتومتری بیانگر افزایش بیان مارکرهای CD133 و CD24 در گروههای حاوی CM و بیان مارکر CD44 در گروه حاوی 50 درصد CM، نسبت به گروه کنترل است. به طور کلی نتایج نشان داد که فرآیند محیط کشت رویی این سلولها تاثیر مثبتی در القاء تمایز سلولهای بنیادی جنینی انسانی به سلولهای پیش ساز کلیه دارد. | ||
| کلیدواژهها | ||
| تمایز؛ سلولهای بنیادی مزانشیمی؛ محیط کشت رویی؛ کلیه | ||
| عنوان مقاله [English] | ||
| Investigating the effect of conditioned medium of mesenchymal stem cells isolated from the kidneys of newborn mice on the differentiation of human embryonic stem cells into kidney progenitor cells | ||
| نویسندگان [English] | ||
| Fatemeh Razeghi1؛ Marjan Nouri2؛ Fattah Sotoodehnejadnematalahi1؛ Ehsan Ehsani3؛ Kazem Parivar1 | ||
| 1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran | ||
| 2Department of Food Science and Technology, Roudehen Branch, Islamic Azad University, Roudehen, Iran | ||
| 3Department of Biology, Roudehen Branch, Islamic Azad University, Roudehen, Iran | ||
| چکیده [English] | ||
| Kidney diseases are an important medical problem worldwide. Since there are limited treatment options for damaged kidneys, stem cell therapy has become an alternative treatment. The aim of present study is to investigate effect of culture medium obtained from mesenchymal stem cells (MSCs-CM) of newborn mice kidneys in percentages of 10, 30 and 50 on differentiation of embryonic stem cells towards kidney epithelial cells. Mesenchymal stem cells were isolated from kidneys of newborn mice, passaged and propagated. Determination of the identity of cells was done using flow cytometry and checking the expression of surface markers CD105, CD29, CD90. In third passage of extracted cells, supernatant culture medium was collected, hESC were cultured and multiplied in the complete culture medium of cells and the differentiation of hES cells into progenitor cells was investigated. The expression of PAX2, ZO1 and CK18 genes was investigated using RT-PCR, expression of CD133, CD24 and CD44 surface markers was investigated using flow cytometry. Flow cytometry results confirmed the mesenchymal nature of the cells. The results of differentiation of hESCs showed that expression of PAX2, ZO1 and CK18 genes increased significantly (p<0.05) in the groups containing supernatant. The results of flow cytometry show an increase in expression of CD133 and CD24 markers in groups containing CM and the expression of CD44 marker in the group containing 50% CM, compared to control group. In general, results showed supernatant culture process of cells has a positive effect on inducing differentiation of human embryonic stem cells into kidney progenitor cells. | ||
| کلیدواژهها [English] | ||
| Distinction, Mesenchymal stem cells, Supernatant culture medium, Kidney | ||
| مراجع | ||
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