بررسی وضعیت بقای سلولی و بیان نشانگرهای عصبی در سلول‌های حاصل از تمایز سلول‌های بنیادی مشتق از بافت چربی در سیستم کشت سه بعدی آلژینات

نوع مقاله : مقاله های پژوهشی

نویسندگان

1 دانشجوی دکتری، گروه علوم تشریحی و بیولوژی مولکولی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی تهران، تهران، ایران

2 استاد، گروه علوم تشریحی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

3 دانشیار، گروه علوم تشریحی، دانشکد‌ه‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

4 دانشجوی دکتری، گروه ژنتیک و بیولوژی مولکولی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

چکیده

مقدمه: مطالعات متعددی برای فراهم کردن تعداد کافی سلول‌های عصبی جهت درمان آسیب‌های نورودژنراتیو انجام شده است. سلول‌های بنیادی جدا شده از بافت چربی، میزان تکثیر و مقاومت زیادی نسبت به آپوپتوز دارند. این سلول‌ها، قابلیت تمایز به دودمان‌های مختلف سلولی از قبیل سلول‌های عصبی را دارند. هیدروژل آلژینات، پلیمر پلی‌ساکاریدی است که دارای ویژگی‌های مناسبی از قبیل زیست‌سازگاری و عدم تحریک سیستم ایمنی می‌باشد. در این مطالعه، تأثیر هیدروژل آلژینات بر میزان بقا و بیان نشانگرهای عصبی سلول‌های حاصل از تمایز سلول‌های بنیادی جدا شده از بافت چربی انسانی بررسی شد.روش‌ها: سلول‌های بنیادی جدا شده از بافت چربی انسانی در محیط کشت عصبی القا و سپس در هیدروژل آلژینات انکپسوله شدند. میزان رشد و تمایز سلول‌ها به ترتیب با استفاده از ارزیابی Doubling time و Real time quantitative reverse transcription polymerase chain reaction (Real time RT-PCR) 7 روز پس از انکپسوله کردن سلول‌ها بررسی شد. آنالیز داده‌ها با استفاده از آزمون One way-ANOVA انجام گردید.یافته‌ها: میزان رشد سلول‌های انکپسوله در هیدروژل آلژینات، کاهش معنی‌داری را نسبت به گروه شاهد نشان داد (001/0 > P). در حالی که میانگین بیان ژن‌های GFAP (Glial fibrillary acidic protein)، Nestin و MAP2 (Microtubule-associated protein 2) در سلول‌های انکپسوله در مقایسه با گروه شاهد به طور معنی‌داری افزایش یافت (001/0 > P).نتیجه‌گیری: اگر چه سرعت تکثیر سلول‌های انکپسوله در هیدروژل آلژینات، در مقایسه با گروه شاهد به طور معنی‌داری کاهش یافت، اما هیدروژل آلژینات می‌تواند باعث افزایش تمایز عصبی سلول‌های انکپسوله شود.

کلیدواژه‌ها

عنوان مقاله [English]

Cell Viability and Neural Markers Expression of Human Adipose-Derived Stem Cells after Neural Induction in Alginate Tridimensional Cell Culture

نویسندگان [English]

  • Zahra Khosravizadeh 1
  • Shahnaz Razavi 2
  • Hamid Bahramian 3
  • Mohammad Kazemi 4
1 PhD Student, Department of Anatomical Sciencesand Molecular Biology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
2 Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
4 PhD Student, Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: Different studies have been done to obtain sufficient number of neuronal cells for treatment of central nervous system injuries. Adipose-derived stem cells have higher proliferation rate and more resistant to apoptosis. These cells can be induced to differentiate along several multilineage cells such as neuron-like cells. Alginate hydrogel is a polysaccharide polymer that has proper properties including biocompatibility with no immunogenicity. In this study, the effect of alginate hydrogel on the cell viability, proliferation rate and neurogenic differentiation of human adipose derived stem cells was evaluated at 14 days after induction.Methods: Adipose-derived stem cells isolated from human, were cultured in neural induction media and seeded into alginate hydrogel. The cell viability and differentiation efficiency of encapsulated cells were evaluated via doubling time and real time quantitative reverse transcription polymerase chain reaction (real time RT-PCR) analysis, respectively. The collected data was analyzed using one-way ANOVA test.Findings: The rate of proliferation of encapsulated cells significantly decreased as compared with control group (P < 0.001). The expression of nestin, glial fibrillary acidic protein (GFAP) and microtubule-associated protein 2 (MAP2) markers significantly decreased as compared with control group (P < 0.001).Conclusion: Altogether cell viability and proliferation rate of encapsulated cells in alginate hydrogel significantly decreased as compared with control group, but alginate hydrogel can promote neural differentiation of adipose-derived stem cells.

کلیدواژه‌ها [English]

  • Alginate hydrogel
  • Adipose-derived stem cells
  • Neurogenic differentiation
  • Proliferation

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مجله دانشکده پزشکی اصفهان
دوره 33، شماره 350 - شماره پیاپی 350
مهر و آبان 1394
صفحه 1491-1505

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سابقه مقاله

  • تاریخ دریافت: 15 مرداد 1393
  • تاریخ بازنگری: 30 فروردین 1403
  • تاریخ پذیرش: 17 فروردین 1401

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