مقایسه‌ی تأثیر پلاسمای غنی از پلاکت (PRP) و سرم جنین گاوی (FBS) بر تکثیر و بقای سلول‌های بنیادی مشتق از چربی در داربست فیبرین

نوع مقاله : مقاله های پژوهشی

نویسندگان

1 دانشجوی کارشناسی ارشد، گروه علوم تشریح، دانشکده‌ی پزشکی و کمیته‌ی تحقیقات دانشجویی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

2 دانشیار، گروه علوم تشریح، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

3 استادیار، گروه علوم تشریح، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

4 کارشناس، گروه علوم تشریح، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

چکیده

مقدمه: امروزه در سلول درمانی و مهندسی بافت، به سلول‌های بنیادی مزانشیمی توجه خاصی می‌شود و جهت رسیدن به تعداد سلول مورد نظر با ویژگی‌های مطلوب، دستیابی به شرایط مناسب کشت ضرورت دارد. از این رو، پژوهش حاضر با هدف مقایسه‌ی تأثیر پلاسمای غنی از پلاکت (PRP یا Platelet rich plasma) و سرم جنین گاوی (FBS یا Fetal bovine serum) بر تکثیر و بقای سلول‌های بنیادی مشتق از چربی (ADSCs یا Adipose derived stem cells) در داربست طبیعی فیبرین انجام شد.روش‌ها: نمونه‌های چربی از سه بیمار، طی جراحی لیپوساکشن به دست آمد. سلول‌های بنیادی از بافت چربی استخراج و کشت شد. سپس سلول‌های پاساژ سوم در داربست فیبرین و مدیوم حاوی PRP 10 درصد (گروه مورد) یا FBS 10 درصد (گروه شاهد) کشت داده شدند. میزان تکثیر و بقای سلول‌ها در روزهای چهارم و هشتم با روش MTT (diphenyl tetrazolium bromide assay 5,2 (yl-2- dimethyl thiazol-4,5)3 و رنگ‌آمیزی با Trypan blue و درصد آپوپتوز سلولی با روش Flow cytometry با استفاده از کیت Annexinᴠ-FITC ارزیابی گردید. اطلاعات جمع‌آوری شد و با استفاده از آزمون t مستقل مورد تجزیه و تحلیل قرار گرفت.یافته‌ها: در روزهای چهارم و هشتم میزان بقا و تکثیر و درصد سلول‌های زنده در گروه مورد نسبت به گروه شاهد به طور معنی‌داری بیشتر و میزان سلول‌ها در مرحله‌ی اولیه‌ی آپوپتوز در گروه مورد نسبت به گروه شاهد به طور معنی‌داری کمتر بود (010/0 > P). میزان سلول‌ها در مرحله‌ی نهایی آپوپتوز، در روز 8 به طور معنی‌داری در گروه مورد کمتر از گروه شاهد بود (001/0 > P).نتیجه‌گیری: تأثیر مثبت PRP بر روند بقا و تکثیر سلول‌های ADSCs در مقایسه با FBS قابل توجه است. 

کلیدواژه‌ها

عنوان مقاله [English]

Comparing the Effect of Platelet Rich Plasma (PRP) and Fetal Bovine Serum (FBS) on Proliferation and Survival of Adipose-Derived Stem Cells in Fibrin Scaffolds

نویسندگان [English]

  • Marzieh Sadeghian 1
  • Batool Hashemibeni 2
  • Mohammad Mardani 2
  • Noushin Amirpoor 3
  • Maryam Aliakbari 4
1 MSc Student, Department of Anatomical Sciences, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran
2 Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Assistant Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences Isfahan, Iran
4 Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: Today, stem cells are the best candidate for cell therapy and tissue engineering. To achieve intended number of cells with optimal characteristics, it is necessary to have appropriate condition of culture. Therefore, in this study, we compared the effects of platelet rich plasma (PRP) and fetal bovine serum (FBS) on proliferation and survival of adipose-derived stem cells (ADSCs) in a natural fibrin scaffold.Methods: Adipose tissue specimens were obtained from 3 patients undergoing liposuction surgery. adipose-derived stem cells were isolated from adipose tissue and cultured and encapsulated in fibrin scaffolds in third passage with culture media containing PRP-10% (experimental group) or FBS-10% (control group). Then, the rate of viability and survival were evaluated using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and Trypan blue staining. Percentage of apoptotic cells was evaluated via flow cytometry using Annexinᴠ-FITC kit on fourth and eighth days. Data were collected and statistical analyzed using t-test.Findings: The rates of viability, survival and percentage of living cells in the experimental group were significantly higher than control group on fourth and eight days (P < 0.010 for all). Percentages of early apoptotic cells in the experimental group were significantly lower than control group on fourth and eight days (P < 0.001 for both) .On the eighth day, percentage of late apoptotic cells in experimental group was significantly lower than control group.Conclusion: This study showed that the positive effect of platelet rich plasma on the survival and proliferation of adipose-derived stem cells in compared with fetal bovine serum was significant.

کلیدواژه‌ها [English]

  • Fibrin
  • Platelet rich plasma
  • Adipose-derived stem cells
  • Proliferation

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مجله دانشکده پزشکی اصفهان
دوره 32، شماره 317 - شماره پیاپی 317
بهمن و اسفند 1393
صفحه 2299-2311

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سابقه مقاله

  • تاریخ دریافت: 07 خرداد 1393
  • تاریخ بازنگری: 08 اردیبهشت 1403
  • تاریخ پذیرش: 17 فروردین 1401

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