یافته های نوین در علوم زیستی

اثر اگزوزوم‌های مشتق از مایع مغزی نخاعی بر تمایز عصبی سلول‌های بنیادی مزانشیمی چربی در داربست هیدروژل آلژینات

نویسندگان

مجتبی چروی 1
جواد بهارآرا 2
پریچهر یغمایی 1
نسیم حیاتی رودباری 1

1 گروه زیست شناسی، دانشکده علوم پایه، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

2 گروه زیست شناسی و مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

https://doi.org/10.52547/nbr.8.4.265
چکیده
امروزه محققین تلاش‌های گسترده‌ای جهت یافتن روش‌های نوین درمانی برای آسیب‌های عصبی انجام داده‌‌اند. در این بین نقش اگزوزوم‌ها در زمینه ارتباطات سلول – سلول به عنوان مکانیسمی نوین قلمداد می‌شود. اگزوزوم‌ها می‌توانند به عنوان عامل تمایزی مناسب عمل کنند. هدف از این پژوهش، بررسی اثر تمایزی اگزوزوم‌های مشتق از مایع مغزی نخاعی بر سلول‌های بنیادی مزانشیمی مشتق از بافت چربی در هیدروژل آلژینات است. در این مطالعه اگزوزوم‌ها به روش اولتراسانتریفوژ از مایع مغزی نخاعی استخراج شده و توسط میکروسکوپ اتمی، میکروسکوپ SEM و تکنیک DLS شناسایی شد. همچنین سلول‌های بنیادی چربی قرار گرفته در هیدروژل آلژینات تحت تیمار با غلظت‌های مختلف اگزوزوم قرار گرفت. بقای سلول‌ها توسط روش‌های MTT و اکردین اورنج اتدیوم بروماید مورد ارزیابی قرار گرفت. بررسی فرایند تمایز سلول‌ها نیز توسط تکنیک ایمنوسیتوشیمی و Real Time – PCR انجام شد. بررسی هویت اگزوزوم‌ها وجود اگزوزوم‌ها با اندازه تقریبی 70 نانومتر را تایید کرد. نتایج بقا سلول‌ها نشان دهنده قابلیت بقا و تکثیر سلول‌ها در طی 14 روز است. همچنین بیان پروتئین‌های MAP2 (پروتئینی همراه با میکروتوبول‌ها) و Nestin (پروتئین فیلامنت بینابینی) با استفاده از روش ایمنوسیتوشیمی مورد تایید قرار گرفت. نتایج حاصل از Real Time – PCR نشان داد سطح بیان ژن MAP2 در طی روز هفتم و چهاردم افزایش و بیان ژن Nestin در مقایسه با گروه کنترل کاهش معنی داری را نشان می‌دهد. این مطالعه نشان داد اگزوزوم‌های استخراج شده از مایع مغزی نخاعی می‌توانند سبب تمایز عصبی سلول‌های بنیادی مزانشیمی چربی در داربست‌های هیدروژلی آلژینات شود.



کلیدواژه‌ها

اولتراسانتریفوژ
ایمنوسیتوشیمی
هیدروژل
مهندسی بافت
وزیکول‌های برون سلولی

عنوان مقاله English

The effect of cerebrospinal fluid-derived exosomes on neural differentiation of adipose mesenchymal stem cells in alginate hydrogel scaffold

نویسندگان English

Mojtaba Cheravi 1
Javad Baharara 2
Parichehreh Yaghmaei 1
Nasim Hayati Roudbari 1
1 Department of Biology, faculty of science, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Department of Biology and Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
چکیده English

Nowadays, researchers have made extensive efforts to find new treatments for nerve damage. Meanwhile, the role of exosomes in cell-cell communication is considered to be a new mechanism. Exosomes can act as suitable differentiating agents. The aim of this study was to investigate the differentiating effect of cerebrospinal fluid-derived exosomes on adipose mesenchymal stem cells in alginate hydrogel. Exosomes were extracted from the cerebrospinal fluid by ultracentrifugation and were then identified by atomic force microscopy (AFM), SEM and DLS technique. In addition, Adipose Mesenchymal Stem cells in alginate hydrogel were treated with different concentrations of exosomes. Cell survival was assessed by MTT and Acridine Orange/Ethidium Bromide methods. Cell differentiation was processed by immunocytochemistry and Real-Time PCR. Examinations confirmed the presence of exosomes with an approximate size of 70 nm. Cell survival results indicate that he ability of cells to survive and proliferate during 14 days. Also, the expression of MAP2 proteins (microtubule-associated protein 2) and Nestin (intermediate filament protein) was confirmed by immunocytochemistry. The results of Real Time - PCR showed that during the seventh and fourteenth days the expression level of MAP2 gene increased and the expression of Nestin gene showed a significant decrease compared to the control group. This study showed that exosomes extracted from cerebrospinal fluid can cause neuronal differentiation of Adipose mesenchymal stem cells in alginate hydrogel scaffolds.





کلیدواژه‌ها English

extracellular vesicles
hydrogel
immunocytochemistry
tissue engineering
ultracentrifugation
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  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405

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