اثرحفاظت انجمادی ملاتونین بر سلول‌های بنیادی اسپرماتوگونی موش نوزاد، افزایش تکثیر و کاهش آپوپتوز

نوع مقاله : Original Article(s)

نویسندگان

1 دانشیار، گروه علوم تشریحی، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی تهران، تهران، ایران

2 دانشجوی ارشد علوم تشریح، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

3 استادیار، گروه علوم تشریح و بیولوژی تولید مثل، دانشکده‌ی پزشکی، دانشگاه علوم پزشکی اصفهان، اصفهان، ایران

10.48305/jims.v43.i838.1488

چکیده

مقاله پژوهشی




مقدمه: با توجه به افزایش میزان ناباروری و نقش کلیدی سلول‌های بنیادی اسپرماتوگونی (SSCS) در شروع اسپرماتوژنژ و حفظ باروری، تلاش برای به حداقل رساندن آسیب‌های ناشی از تکنیک‎های نگه‎داری طولانی مدت SSCs یکی از مباحث حائز اهمیت در تحقیقات پایه و بالینی محسوب می‌شود. بنابراین این مطالعه به منظور بررسی اثر ملاتونین بر سلول‎های بنیادی اسپرماتوگونی نوزاد موش طی فرایند انجماد و ذوب طراحی شد.
روش‌ها: سلول‌های بنیادی اسپرماتوگونی از نوزادان موش‌ نر 5-3 روزه نژاد BALB/C بعد از هضم آنزیمی جداسازی شده و با نشانگر  ID4توسط تکنیک فلوسایتومتری تایید ماهیت شدند. بعد از کشت، سلول‌ها در دو گروه کنترل (محیط پایه انجمادی) و گروه 100 میکرومولار ملاتونین در محیط انجمادی پایه به مدت یک ماه فریز شدند. پس از فرایند انجماد– ذوب، میزان بیان پروتئین آپوپتوزی Caspase-3  و بیان پروتئین تکثیری Gfra1  در سلول‌ها مورد ارزیابی قرار گرفت. سطح معنی‌داری 0/05 ≥ P در نظر گرفته شده است.
یافته‌ها: پس از فرایند انجماد- ذوب، ملاتونین سبب کاهش معنادار بیان پروتئین آپوپتوزی Caspase-3  شد. همچنین افزایش بیان پروتئین GFRa1 در حضور ملاتونین به صورت معنی‌داری نسبت به گروه کنترل گزارش شد.
نتیجه‌گیری: نتایج حاصل از این مطالعه نشان داد که ملاتونین، به عنوان یک عامل محافظ انجماد از طریق کاهش آپوپتوز و افزایش تکثیر سلول‎های بنیادی اسپرماتوگونی می‌تواند برای انجماد و نگه‎داری طولانی مدت سلول‎ها و درمان ناباروری در کلینک گزینه مناسبی ‎باشد.

تازه های تحقیق

عطیم هدایت پور:  PubMed ,Google Scholar 

مریم خانه زاد:  Google Scholar 

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

The Cryopreservation Effect of Melatonin on Neonatal Mouse Spermatogonial Stem Cells, Increasing Proliferation and Decreasing Apoptosis

نویسندگان [English]

  • Azim Hedayapour 1
  • Reihaneh Ebneyamin 2
  • Maryam Khanehzad 3
1 Associate Professor, Department of Anatomical Sciences, Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
2 MSc Student, Department of Anatomical Sciences and Reproductive Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
3 Assistant Professor, Department of Anatomical Sciences and Reproductive Biology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
چکیده [English]

Background: Considering the increasing infertility rate and the key role of spermatogonia stem cells (SSCS) in initiating spermatogenesis and maintaining fertility, trying to minimize the damage caused by long-term storage techniques of SSCs is considered as one of the important topics in basic research and clinic. Therefore, this study was designed to investigate the effect of melatonin on neonatal mouse spermatogonial stem cells following freezing and thawing.
Methods: Spermatogonial stem cells were isolated from neonatal mouse 3-5 days old BALB / C after enzymatic digestion and identified with IDA marker by flow cytometry. After culture, the cells were frozen in two groups:1) control (basal cryopreservation) and 2) 100 μM melatonin in basal cryopreservation for one month. After freezing-thawing process, the expression level of the apoptotic protein Caspase-3 and the expression of the proliferative protein Gfra1in the cells were evaluated. The significance level was considered to be P ≤ 0.05.
Findings: After freezing-thawing process, melatonin significantly reduced the expression of the apoptotic protein Caspase-3. Also, a significant increase in the expression of Gfra1protein was reported in the presence of melatonin compared to the control group.
Conclusion: The results of this study indicate that melatonin as a cryoprotectant by reducing apoptosis and increasing proliferation of spermatogonial stem cells can be a suitable option for cryopreservation and long-term storage of cells and infertility treatment in the clinic.

کلیدواژه‌ها [English]

  • Spermatogonial Progenitor Cells
  • Melatonin
  • Cryoprotective Agent

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مجله دانشکده پزشکی اصفهان
دوره 43، شماره 838
هفته 2، دی
آذر و دی 1404
صفحه 1488-1496

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  • تاریخ دریافت: 28 آذر 1404
  • تاریخ پذیرش: 28 دی 1404

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