تاثیر فاکتور سلول بنیادی و رتینوئیدها در تمایز سلول های اسپرماتوگونی بافت بیضه رت بالغ مدل آزواسپرمی انسدادی

نوع مقاله: مقاله پژوهشی

نویسندگان

1 گروه زیست شناسی، واحد آیت ا... آملی، دانشگاه آزاد اسلامی، آمل، ایران

2 گروه علوم تشریح، مرکز تحقیقات بهداشت باروری و ناباروری، پژوهشکده سلامت، دانشگاه علوم پزشکی بابل، ایران

3 گروه مامایی، مرکز تحقیقات باروری بهداشت، واحد ساری، دانشگاه آزاد اسلامی، ساری، ایران

4 گروه فارماکولوژی، مرکز تحقیقات بیولوژی سلولی و مولکولی، پژوهشکده سلامت، دانشگاه علوم پزشکی بابل، ایران

چکیده

رتینوئیدها و فاکتور سلول بنیادی نقش مهمی در تکثیر، بقا و تمایز سلول­های بنیادی به اسپرم ایفا می­کنند. لذا در این مطالعه اثر فاکتور سلول بنیادی و رتینوئیدها بر تکثیر و تمایز سلول­های اسپرماتوگونی در لوله­های اسپرم­ساز بیضه رت بالغ مدل آزواسپرمی انسدادی با استفاده از سیستم کشت بافت مورد ارزیابی قرار گرفته است. قطعات بافت بیضه در محیط کشت مکمل با رتینوئیدها یا فاکتور سلول بنیادی و یا ترکیبی از هر دو در شرایط دمای 5/34 درجه سانتی­گراد و دی اکسید کربن 5 درصد به مدت 25، 30 و 35 روز کشت داده شدند. سپس مقاطع بافتی تهیه شده در زمان­های مذکور به وسیله رنگ­آمیزی پاس و هماتوکسیلین مورد ارزیابی مورفولوژیکی قرار گرفتند. شمارش سلولی در واحد سطح با استفاده از نرم­افزارImageJ  و تجزیه و تحلیل آماری، با استفاده از آنالیز واریانس یک طرفه، ANOVA و آزمون تعقیبی توکی انجام شد. تعداد سلول­های اسپرماتوگونی، اسپرماتوسیت و لایدیگ در روز 25، 30 و 35 کشت و همچنین تعداد سلول اسپرماتید گرد و دراز در روز 35 کشت در بافت بیضه آزواسپرمی انسدادی در مقایسه با گروه­های تجربی و کنترل افزایش معنی­دار داشتند (001/0>p ). بررسی هیستولوژیک بافت بیضه در شرایط مختلف کشت، تغییرات پاتولوژیک را در سلول­های سازنده اپی­تلیوم نشان داد. رتینوئیدها در روز 35 کشت در مقایسه با فاکتورهای دیگر در حفظ و تکثیر رده­های سلولی فرایند اسپرماتوژنز نقش مؤثرتری داشتند. از آنجائیکه این فاکتورها به تنهایی عامل القا تکثیر و تمایز نیستند، بنابراین بهینه سازی محیط کشت جهت دستیابی به اسپرم بالغ پیشنهاد می­شود.

کلیدواژه‌ها


عنوان مقاله [English]

The Effect of Stem Cell Factor and Retinoids in Spermatogonial Cell Differentiation of Testicular Tissue of Obstructive Azoospermia Model Adult Rats

نویسندگان [English]

  • Mahnaz Nasimi 1
  • Esmaeil Fattahi 1
  • Sayed Gholamali Jorsaraei 2
  • Maryam Gholamitabar Tabari 3
  • Ebrahim Zabihi Neyshabouri 4
1 Department of Biology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran
2 2- Department of Anatomical Sciences, Infertility and Reproductive Health Research Center, Research Institute for Health, Babol University of Medical Sciences, Babol, Iran
3 Department of Midwifery, Research center for Fertility Health, Sari Branch, Islamic Azad University, Sari, Iran
4 Department of Pharmacology, Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran
چکیده [English]

Retinoids and stem cell factor play an important role in the stem cells proliferation, survival, and differentiation into sperm. Therefore, in this study, the effect of stem cell factor and retinoids on the proliferation and differentiation of spermatogonial cells in testis seminiferous tubules of obstructive azoospermia model adult rats was investigated using the tissue culture system. The testicular tissue fragments were cultured in the culture medium supplement with retinoids or stem cell factor, or a combination of both at the temperature of 34.5 °C and 5% CO2 for 25, 30, and 35 days. Then, the stained tissue sections prepared at the mentioned times were evaluated using PAS and hematoxylin staining methods. The cell counting per unit area was done using ImageJ software, and the statistical analysis was performed by one-way ANOVA and Tukey's post hoc test. The number of spermatogonia, spermatocyte, and Leydig cells increased significantly on day 25, 30 and 35 of culture; additionally, the number of round and elongated spermatid increased significantly on day 35 of culture in the testicular tissue of the obstructive azoospermia compared to the experimental and control groups (p < 0.001). Histological examination of testicular tissue in different culture conditions revealed pathological changes in the epithelialmaking cells. Retinoids played a more effective role on day 35 of culture compared to other factors in maintaining and proliferation of cell lines of the spermatogenesis process. As these factors are not alone the cause of proliferation and differentiation induction, therefore optimizing the culture medium is recommended to achieve adult sperm.

کلیدواژه‌ها [English]

  • Retinoids
  • Stem Cell Factor
  • tissue culture
  • Obstructive Azoospermia
  • Rat
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